Clinica Chimica Acta 330 (2003) 1 – 30

www.elsevier.com/locate/clinchim

Review
Capillary electrophoresis and its application in the
clinical laboratory
John R. Petersen*, Anthony O. Okorodudu, Amin Mohammad, Deborah A. Payne
Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA
Received 8 July 2002; received in revised form 6 December 2002; accepted 17 December 2002

Abstract

Over the past 10 years, capillary electrophoresis (CE) is an analytical tool that has shown great promise in replacing many
conventional clinical laboratory methods, especially electrophoresis and high performance liquid chromatography (HPLC). The
main attraction of CE was that it was fast, used small amounts of sample and reagents, and was extremely versatile, being able to
separate large and small analytes, both neutral and charged. Because of this versatility, numerous methods for clinically relevant
analytes have been developed. However, with the exception of the molecular diagnostic and forensic laboratories CE has not had
a major impact. A possible reason is that CE is still perceived as requiring above-average technical expertise, precluding its use in
a laboratory workforce that is less technically adept. With the introduction of multicapillary instruments that are more automated,
less technique-dependent, in addition to the availability of commercial and cost effective test kit methods, CE may yet be
accepted as a instrument routinely used in the clinical laboratories. Thus, this review will focus on the areas where CE shows the
most potential to have the greatest impact on the clinical laboratory. These include analysis of proteins found in serum, urine, CSF
and body fluids, immunosubstraction electrophoresis, hemoglobin variants, lipoproteins, carbohydrate-deficient transferrin
(CDT), forensic and therapeutic drug screening, and molecular diagnostics.
D 2003 Elsevier Science B.V. All rights reserved.

Keywords: Capillary electrophoresis; Clinical laboratory; Clinical applications; Molecular diagnostics

Abbreviations: AGE, agarose gel electrophoresis; CAE, cellu-

lose acetate; CE, capillary electrophoresis; CZE, capillary zone
electrophoresis; CGE, capillary gel electrophoresis; CIEF, capillary
isoelectric focusing; MEKC, micellar electrokinetic chromatogra-
phy; CITP, capillary isotachophoresis; CE-IS, capillary electro-
phoresis, immunosubstraction; CSF, cerebrospinal fluid, immuno-
electrophoresis; CDT, carbohydrate-deficient transferring; HPCEC,
high performance cation-exchange chromatography; IEP, immuno-
electrophoresis; IFE, immunofixation electrophoresis; LOH, loss of
heterozygosity; MC, monoclonal component; SNP, single nucleotide
polymorphisms; STR, short tandem repeat; TDM, therapeutic drug
monitoring.
* Corresponding author. Tel.: +1-409-772-1350; fax: +1-409-
772-9231.
E-mail address: jrpeters@utmb.edu (J.R. Petersen).
1. Introduction
Clinical laboratories are under tremendous pressure
to develop assays that are accurate, precise, fast, cap-
able of being automated, and yet, inexpensive. For
electrophoresis, this has been difficult. However, by
using capillary electrophoresis (CE) many, if not all, of
these goals can potentially be attained. This is because
CE, even though based on the movement of molecules
in an electric field, is not restricted to areas classically
assigned to electrophoresis, namely separation of large
molecules based on size or charge. It can also separate
molecules that have low molecular weight, in addition

0009-8981/03/$ - see front matter D 2003 Elsevier Science B.V. All rights reserved.
doi:10.1016/S0009-8981(03)00006-8
2 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
to neutral compounds, such as steroids. The commonly
used modes of CE, which in many instances can be
accessed by simply altering the buffer composition, are
capillary zone electrophoresis (CZE), gel electropho-
resis (CGE), capillary isoelectric focusing (CIEF),
micellar electrokinetic chromatography (MEKC), and
capillary isotachophoresis (CITP). CZE, where a capil-
lary is filled with a relatively low viscosity buffer, is
the simplest form of CE. The analytes of interest
migrate from one end of the capillary to the other with
velocities determined by the charge to mass ratio to
form discrete peaks (zones). CGE separates analytes,
such as DNA or proteins, based on molecular weight.
A variety of compounds, such as bis-polyacrylamide,
agarose, or methylcellulose, have been used to act as
molecular sieve to cause separation. MEKC takes
advantage of a property of surfactants, such as sodium
dodecyl sulfate (SDS), known as the critical micelle
concentration where micelles (aggregates) are formed
once the surfactant reaches a specific concentration.
By taking advantage of the differential distribution of
various analytes in the micelles separation of neutral
and charged analytes is possible. CIEF is similar to
classical isoelectric focusing where peptides and pro-
teins are separated on the basis of isoelectric point (pI)
by generating a pH gradient using ampholytes. The
difference is that instead of creating a pH gradient in a
gel it is created inside a capillary. CITP is similar to
CZE, except that a discontinuous buffer (a combina-
tion of two buffers that have different mobilities) is
used causing the analytes of interest to be concentrated
in zones between the leading and terminating constit-
uents. Enhanced separation can be achieved by using
spacer compounds, such as various amino acids. The
large number of methods and instruments, such as the
traditional chromatographic techniques of gel electro-
phoresis, GC, and HPLC, that potentially can be
replaced by CE along with the high separation effi-
ciency (high number of theoretical plates) make CE an
attractive technique in the clinical laboratories.
Over the last 5 years, a number of review articles
[1– 12] and books [13,14] have been written on the
use of CE in the clinical laboratory, however, with the
exception of molecular diagnostics CE still has not
had a major impact in the clinical laboratory. Al-
though there are over 900 tests that can be performed
in the clinical laboratory to help in the assessment of a
patients health status it is unreasonable to think that
CE can be only used in more than a small fraction of
these tests. Thus, this review will focus on the areas
where CE shows the potential to have the greatest
impact in the clinical laboratory and include analysis
of various serum and body fluid proteins, forensic and
therapeutic drug screening, and molecular diagnostics.
Immunoassay, which shows future promise, especially
with the advent of chip technology, will not be
covered.
2. Applications of capillary electrophoresis in
clinical settings:
2.1. Protein analysis
2.1.1. Serum protein
Currently, the CE method that has had the most
extensive clinical validation is in the analysis of serum
proteins. The first indication that CE could be used in
the separation of serum proteins into the classic six
bands, e.g. albumin, a-1 globulin, a-2 globulin, h-1
and h-2 globulins, and g globulin, was reported by
Jorgenson and Lukas [15]. Although variations in these
protein fractions can be correlated with a patient’s
health status it is usually the changes that occur in h
and g fractions that are of the most clinical interest. It is
in these fractions that monoclonal proteins are nor-
mally found and whose detection is an important part of
the laboratory evaluation of patients with lymphopro-
liferative diseases. Large monoclonal bands are usually
present in patients with multiple myeloma or Walden-
ström’s disease; however, lower concentrations may be
seen in a variety of other diseases such as light chain
disease, leukemia, lymphoma, amyloidosis, or mono-
clonal gammopathy of undetermined significance
(MGUS). Patients that fall into the MGUS category
should have annual follow-ups because the chance that
these patients will develop myeloma or other lympho-
proliferative diseases is increased [16]. Classically,
screening serum has been done by either cellulose
acetate (CAE) or agarose gel (AGE) electrophoresis.
While the reagents for both of these methods are
relatively inexpensive, these methods require a signifi-
cant amount of labor. Considerable time is spent by
technologists in applying the samples, fixing, staining,
washing the gels to remove excess stain, and perform-
ing the densitometric scans. Although there has been
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
increased automation, classical electrophoresis meth-
ods still require a significant amount of labor and it is
for this reason that various groups have evaluated CE
with the hope that the labor component could be
reduced.
Although numerous research groups [17 – 19], in-
cluding one from the industry sector [20], have looked
at ways to improve on the initial method of Jorgenson
and Lukas, none looked at large numbers of patients to
fully evaluate the method as a replacement for AGE or
CAE. It was not until Jenkins and Guerin [21 – 23],
who used a single capillary CE instrument, was CE
compared to AGE with large numbers of clinical sam-
ples. In these reports, a total of 6500 samples, of which
1357 contained a monoclonal component (MC), were
compared. They were able to detect four IgA and one
IgG MC that were not detected by AGE. Conversely,
they reported that CE missed five IgM and three IgG
MCs. However, by increasing the ionic strength and
pH of the run buffer they were able to detect these MC
indicating the importance of method validation with
known samples. CE also missed a 1 g/l light chain
band that was visible in AGE although in 11 other
cases free light chain was detected by both CE and
AGE. Although eight mini-MCs co-migrating with C3
were not detected by CE, they reported that most mini-
MCs in the range of 0.5– 1 g/l were usually present as
an irregularity in the gamma region. Due to potential
interferences, such as elevated C-reactive protein or
oligoclonal banding due to an acute inflammatory
response identification of an irregularity as a mini-
MC required careful interpretation [24]. They also
reported a good correlation (r = 0.96) between CE
and AGE in estimating the monoclonal concentrations
ranging from 1 to 71 g/l. In a prospective study,
Katzmann et al. [25] used immunotyping to identify
the presence or absence of an MC in 1518 clinical
samples. They found that CE was more sensitive than
AGE (94.9% vs. 90.7%) with similar specificity
(98.6% vs. 98.9% for AGE). This was confirmed by
Bossuyt et al. [26] using 58 specimens previously
identified as containing an MC. Using immunofixa-
tion as the gold standard CE was able to detect an MC
in 95% of the known samples. The same group found
the same results in a prospective study of 1692 pa-
tients that were being screened for the presence of an
MC or as follow-up of a known MC [27]. More re-
cently, Jonsson et al. [28] developed computer algo-
3
rithms to detect MCs in the h and g regions. Using
these algorithms, only one MC (concentration 1 g/l)
out of 95 was not identified for a sensitivity of 98.9%
while correctly identifying 604 of 607 samples that
did not have an MC for a specificity of 99.5%. They
were also able to correctly identify an MC due to
circulating light chain in serum samples from five of
nine patients with Bence – Jones proteinuria even
though light chain was not visible in two of these
serum samples by AGE.
In the initial evaluation of the Beckman Paragon
CZE 2000 (Beckman Coulter, Fullerton, CA, USA),
the first commercial CE developed exclusively for
clinical purposes, Jolliff and Blessum [29] compared
the precision of CE and AGE. They found that the
within-capillary precision for CE was 0.6 – 3.3%
(N = 10) and the between-capillary precision was
1.8– 4.5% (N = 120), compared to an AGE precision
of 3.8 – 8.0% (N = 120) for the classic five serum
protein bands. This confirmed a previous study by
Winen and van Dieijen-Vesser [32] who found a
within- and between-capillary precision of < 3.1% for
all protein bands. In addition, Jolliff and Blessum
evaluated the ability of CE and AGE to discriminate
between various disease states in 240 patient samples.
They found that, although the concentrations of the
various protein fractions were not identical, there was a
96% concordance between the two methods in the
discrimination of various disease states involving a
variety of dysproteinemias, such as hypogammaglobu-
linemia, acute and chronic inflammation, nephrosis,
cirrhosis, poly- and monoclonal gammopathies. A
normal serum protein electrophoregram using the
Beckman Paragon CZE 2000 is shown in Fig. 1.
Bossuyt et al. [30] reached similar conclusions when
they studied 524 patient samples covering a similar
population of dysproteinemias. They found a poor
correlation between the concentrations of each protein
fraction determined by CE and AGE (r < 0.61 for all
protein fractions), although there was a reasonable
correlation with CAE (r>0.85 for all fractions except
the h globulin which was 0.67). These differences can
be traced to the use of dye binding to detect the various
proteins in AGE and CAE vs. CE’s use of the amide
bond absorbance at 200 – 214 nm. This will affect the
quantification but will not change the clinical interpre-
tation. Petrini et al. [31] established normal ranges for
CE using 167 samples that were normal by CAE and
4 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30

Fig. 1. Serum protein electrophoresis using the Beckman Paragon CZER 2000. Normal serum protein electrophoregram seen using the Beckman
Paragon CZER. All parameters are preset by manufacture. Separation Conditions: capillary length is 20 cm (18 cm to detector)  25 Am (I.D.);
detection was at 214 nm; temperature, 24 jC; a proprietary buffer is used at 10.5 kV for 4 min; injection by vacuum for 1 s. Superimposed on the
electrophoregram is where the major components of the various protein bands would be expected to be found. The figure was kindly supplied by
Beckman Coulter.

then verified the ranges using one thousand normal and
abnormal serum samples. These studies have been
confirmed by other groups in comparing CE with
AGE [32 – 38]. Recently, Bossuyt et al. [39] expanded
on these normal range studies by establishing pediatric
serum protein reference ranges using a small number of
children (151 males and 44 females, ages 1 – 16 years).
Although there were differences between CE and AGE/
CAE, in particular the reference ranges (see Table 1);
CE was found to be an efficient, reproducible method
for the separation of serum proteins that is able to iden-
tify abnormalities that are not detected by either AGE
or CAE. In addition, the technical skills required for the
newer CE instruments, such as Beckman Coulter’s
Paragon CZE 2000k or Sebia’s CAPILLARYSk,
are less than those required for AGE/CAE making
CE is a viable alternative to conventional electropho-
resis, especially when large numbers of samples are
submitted for serum protein analysis.
The ability to identify whether a pleural fluid is a
transudate or exudate can affect patient care. Inaccu-
rate classification can lead to unnecessary invasive
procedures. To overcome this problem, a set of criteria
was developed by Light et al. [56] to help identify
exudates. While this criteria is sensitive (>95%), it is
not specific (f 70%). Recently, CE has been used to
analyze body fluids, pleural and ascetic, from a small
number of patients (N = 47) using methods developed
for serum protein separations [55]. By using CE to
calculate the a2-macroglobulin/albumin ratio, the
authors reported that CE could identify exudates with
a sensitivity of 81% and a specificity of 91%. Combin-
ing the Light criteria with the a2-macroglobulin/albu-
min ratio obtained by CE, all exudates were correctly
identified (sensitivity—100%); however, a small num-
ber of transudates (2 out of 11) were incorrectly
identified. Although evaluation of additional samples
is needed to confirm the initial results of Claeys et al.,
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30 5

Table 1
Reference ranges (% total protein content) for the five serum protein fraction by capillary electrophoresis. Comparison of representative
literature results with agarose gel electrophoresis
Author Reference Age Albumin Alpha 1 Alpha 2 Beta Gamma
(years) (%) (%) (%) (%) (%)
Beckman Coulter Package Adult 49.7 – 64.4 4.8 – 10.1 8.5 – 15.1 7.8 – 13.1 10.5 – 19.5
(Paragon CZE 2000) Insert
Sebia (Capillarys CE) Package Adult 56 – 66 3.6 – 6.0 6 – 10 9 – 14.5 11 – 19
Insert
Jolliff and Blessum [29] Adult 52.0 – 67.0 3.8 – 8.3 5.8 – 12.4 9.8 – 13.0 9.3 – 20.4
(Paragon CZE 2000)
Bossnyt et al. [30] Adult M 41.7 – 52.3 2.6 – 4.5 3.4 – 6.4 5.8 – 9.5 5.3 – 13.2
(Paragon CZE 2000) F 37.4 – 49.8 2.6 – 5.1 3.9 – 6.4 5.5 – 8.7 9.6 – 20.5
Petrini, et al. [31] Adult 53.0 – 67.6 3.3 – 8.4 5.0 – 11.0 8.1 – 13.3
(Paragon CZE 2000)
Bossnyt et al. (Pediatric) [39] Age 1 – 2 54.7 – 70.4 4.2 – 8.5 7.0 – 15.6 7.5 – 11.6 4.7 – 16.0
(Paragon CZE 2000) Age 3 – 4 53.9 – 70.4 4.8 – 8.1 7.6 – 15.2 7.4 – 11.5 7.1 – 17.8
Age 5 – 9 52.6 – 66.3 4.2 – 7.6 7.4 – 13.5 7.9 – 11.3 8.5 – 18.7
Age 10 – 14 54.1 – 69.1 4.4 – 8.0 6.8 – 11.4 8.5 – 12.9 8.8 – 17.6
Helena SPIFE 2000 Package Adult 46.6 – 62.6 1.7 – 4.1 5.9 – 13.5 10.9 – 18.9 11.6 – 24.4
Insert
Sebia Hydragel Package Adult 59.7 – 70.6 1.4 – 2.7 7.2 – 11.1 8.0 – 14.7 8.4 – 16.3
Insert
Beckman Paragon Package Adult 54.6 – 68.5 3.6 – 7.8 5.2 – 10.7 8.5 – 13.7 9.7 – 19.2
Insert

the combination of Light’s criteria in conjunction with 2.1.2. Immunosubstraction electrophoresis

the a2-macroglobulin/albumin ratio using CE should
aid in the identification of transudates enhancing
patient care.
A possible exception to the usefulness of CE to
identify serum proteins abnormalities is its apparent
inability to detect an a-1 antitrypsin deficiency, spe-
cifically when the SS, SZ, and MS phenotypes are
present [40], although the ZZ phenotype is detectable
[30,40]. Although only one report, Gonzalez-Sagrado
et al. indicated that if the concentration of the a-1
globulin fraction by CE was less than 4 g/l, then
quantitation of a-1 antitrypsin is required. Perhaps,
separate quantification of the a-1 acid glycoprotein
and a-1 antitrypsin areas, which are distinguishable by
CE (see Fig. 1), would eliminate this problem. How-
ever, until this issue can be resolved, AGE—and not
CE—should be used as a screening method if a
deficiency in a-1 antitrypsin is suspected. Even though
CE should not be used to screen for a-1 antitrypsin
deficiency, it has been shown to be useful in the
phenotyping of a-1 antitrypsin and thus could be used
to identify a-1 antitrypsin variants associated with
clinical disease [41].
In addition to detection, classification or typing of
an MC is important in the long-term follow-up of
patients [16]. For many of these patients, quantifica-
tion and typing is used as a marker to help determine
when malignancy has occurred or to monitor therapy.
The most common methods to type an MC are by
immunoelectrophoresis (IEP) or immunofixation elec-
trophoresis (IFE). Although IFE is more sensitive than
IEP [42 – 44], both are technically demanding and
require a significant amount of labor. At present,
neither of the classical forms of IFE nor IEP is possible
by CE. However, a CE compatible method that can be
used to type MC has been developed by modifying a
procedure described by Aguzzi and Poggi [45]. This
technique is called immunosubstraction electrophore-
sis or CE-IS. The method involves the removal of
specific proteins from a sample using beads coated
with anti-heavy chain (IgG, IgM, IgA) or anti-light
chain (n or E) antibodies before separation by CE [46].
Identification is based upon which of the immunosor-
bants removes the MC (Fig. 2).
Currently, the only commercial clinical instrument
that uses CE-IS to type an MC is the Paragon CZE
6 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30

Fig. 2. CE-IS of a monoclonal IgM-n using the Beckman Paragon CZER 2000. CE immunosubtraction using the Beckman Paragon CZER. All
parameters are preset by manufacture. Separation Conditions: capillary length is 20 cm (18 cm to detector)  25 Am; temperature, 24 jC;
a proprietary buffer was used at 9.5 kV for 5 min; injection by vacuum for 1 s. The serum protein electropherogram (SPE) shows a monoclonal
peak in the beta region. After treatment of the serum is treated with beads coated with the various antibodies (IgG, IgM, IgA, n, E) the
monoclonal peak is removed using beads coated with anti-IgM and anti-n. The monoclonal protein is thus an IgM-n. The figure was kindly
supplied by Beckman Coulter.

2000. In the first published study using this instrument,
Jolliff and Blessum evaluated the ability of CE-IS to
correctly type an MC and found that if an MC was
detectable (>0.5g/l for IgG and >0.75 g/l for IgA or
IgM), the results of CE-IS were identical with IFE
[29]. This was confirmed by Katzmann et al. [25], who
also found that when an abnormality was visible in the
electropherogram, CE-IS was able to correctly type the
MC. The same group of researchers also found that
CE-IS is easier to perform than IFE, and was extremely
helpful in the interpretation of abnormalities that are
difficult to analyze using AGE/IFE [47]. In a smaller
study on 58 patients with an MC previously identified
by IFE/IEP, Bossuyt et al. [48] found that the CE-IS is
superior to CAE and equivalent to AGE in the iden-
tification of an MC. They indicated that increased
automation and precision make CE-IS an acceptable
replacement for the currently used IEF methods to
detect, quantify, and type MC. They did note, however,
that CE-IS, like CAE and AGE, may miss a low-
concentration MC due to an MGUS, light chain dis-
ease, some lymphomas, etc., and suggested that if an
MC is clinically suspected, a complimentary, more
sensitive method, such as IFE, should be used. Similar
conclusions were also reached by Bienvenu et al. [49]
and Henskens et al. [50]. Litwin et al. [51], however,
were not convinced of the utility of CE-IS. In their
study, they first compared the ability of CE and AGE
to detect the presence of an MC and found, like other
researchers, that CE was more sensitive than AGE.
However, the second part of the study, evaluating the
ability to correctly interpret CE-IS results, found that
four individuals were able to correctly identify and
type only 60 – 75% of the MC identified by IFE.
Although this is unacceptable, if the MC was prom-
inent and discrete all MC were properly identified.
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
Interpretation of the subtle differences in the electro-
phoresis before and after CE-IS was difficult only
when the MC was small and/or superimposed on a
polyclonal background. As with any new technology,
this problem may be resolved by appropriate, focused
training on how to interpret the electropherograms or
use of computer algorithms, such as the version
developed by Jonsson et al. [28] to help interpret
electrophorograms before and after IS. Overall, CE-
IS is an excellent alternative to IFE for the great
majority of MC evaluations. However, if a question
remains after CE-IS then IFE should be used to
confirm the interpretation.
It is also possible to use CE to detect cryoglobulins
and, although typing of cryoglobulins using CE-IS has
not been published, it would obviously be the next step
in the evaluation. Like AGE, detection by CE is
sometimes difficult due to precipitation of the cryo-
globulin at temperatures lower than 37 jC. However,
visual inspection of the electropherogram along with
clinical information will usually indicate the presence
of a serum abnormality that requires further investiga-
tion [33,52,53]. The advantage that CE has over AGE
is that it is able to detect an MC in the cryoglobulin
precipitate even when only low levels of protein are
available. If quantification is desired precipitation of
the cryoglobulin followed by redissolving, the precip-
itate before analysis is usually required. However, if
the cryoglobulin concentration is < 4 g/l, underesti-
mation can occur [54]. Thus, CE along with the future
use of CE-IS, will give physicians additional informa-
tion about a cryoglobulin faster and in a more cost-
effective manner.
2.1.3. Urine proteins
Increased excretion of protein in urine (proteinuria)
is one of the most common abnormalities seen in the
clinical laboratory and can be caused by a number of
pathologic conditions affecting the kidney and urinary
tract [57,58]. Characterization of these proteins by
electrophoresis can be helpful in determining the cause
of increased urinary protein, whether tubular or glo-
merular. Classically, analysis has been by carried out
AGE or CAE, although more recently CE has been
successfully used to analyze urine proteins pattern.
One of the problems with analyzing urine proteins
by CE is that many compounds present in urine can
absorb at the same wavelength (200 or 214 nm) that is
7
used to detect proteins, causing interpretation prob-
lems. To overcome this problem, Friedberg and Sha-
habe [59] used ethanol precipitation or low molecular
weight cut-off filters (>15,000) along with washing,
to concentrate urine proteins. They found good agree-
ment between CE and AGE for selective and non-
selective proteinuria and the detection of MC in urine.
In a separate paper, Jenkins [60] successfully used CE
to separate and detect Bence –Jones proteinuria (range
0.04 – 9.7g/l), intact immunoglobulins, and Tamm
Horsfall proteins in unconcentrated urine samples.
More recently, Kolios et al. [61] used CE to evaluate
unconcentrated urine from patients with glomerular,
tubular, mixed tubular and glomerular, and overflow
proteinuria. They found that as long as the protein
concentration is slightly above normal (>150 – 200
mg/l), the analysis correlated very well with AGE.
Both CE and AGE are able to distinguish tubular
from glomerular proteinuria and detect the presence
of free light chain. The advantage that CE holds over
AGE is that concentration of the urine is not neces-
sary, making CE faster and more cost-effective. In
addition, although low molecular weight (< 10,000)
substances have yet to be shown to have clinical
significance, the ability to detect these substances in
unconcentrated urine by CE may help give a more
complete picture of the pathologic process occurring
in the kidney.
2.1.4. Cerebrospinal fluid proteins
The analysis of cerebrospinal fluid (CSF) proteins
can be useful in the diagnosis and management of a
variety of neurological diseases including conditions
that cause immune responses, destructive brain dis-
eases, or a breakdown in the blood – brain barrier. The
immune process may also produce a polyclonal or,
perhaps more importantly, oligoclonal bands in the
gamma region, which if combined with the IgG index
can be helpful in making a diagnosis of multiple
sclerosis in >95% of the cases [62]. Oligoclonal band
may also be present in patients with a variety of other
neurological conditions [63 – 65]. At present, the
method routinely used in the clinical laboratory for
the detection of protein profiles in CSF is AGE, which
is labor-intensive and requires concentration of the
CSF to visualize the protein bands.
Crowdrey et al. [66] analyzed unconcentrated CSF
from 30 random patients and were able to separate
8 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
between 20 and 25 peaks. Differences were noted in
the electrophorograms; however, no attempt was made
to correlate these differences with disease states nor
was the method able to distinguish the fine features in
the gamma region. Hiraoka et al. [67,68] used capillary
gel electrophoresis to analyze proteins present in CSF.
They were able to separate both high and low molec-
ular weight proteins and found that an index based on
the serum and CSF albumin and a2-macroglobulin was
potentially useful in evaluating the blood –brain bar-
rier in patients with neurological disorders. Manabe et
al. [69] used capillary isoelectric focusing (CIEF) to
separate proteins in unconcentrated CSF and, although
dialysis was required, they were able to identify
proteins in the IgG region that were not obvious in
the serum of the same patient. Sanders et al. [70]
modified a method used to separate serum proteins for
the analysis of CSF. This method gave a concordance
of 87% when compared to high-resolution AGE in the
detection of oligoclonal banding. When trying to use
unconcentrated CSF, however, they found that a 3- to
6-fold increase in the injection volume was required to
detect oligoclonal banding resulting in a loss of reso-
lution. Sanders et al. speculated that by using extended
path length cells (bubble or Z-cells) that give a 5- to
10-fold increase in sensitivity without a corresponding
loss of resolution, detection of oligoclonal bands in
unconcentrated CSF may be possible. One interesting
feature of CE is its ability to calculate a CSF Ig index.
The concentration of the albumin and immunoglobulin
region can be estimated by using CE to separate both
serum and CSF proteins. Substituting these values into
the classic CSF IgG index equation [71] gives a CSF Ig
Index. A good correlation (r = 0.934) was found when
they were compared to the results obtained using
nephelometry with only 2 discordant results out of
24 [70]. A side benefit of using CE to detect oligoclo-
nal banding is this ability to determine the presence or
absence of oligoclonal bands in addition to calculating
a CSF Ig index using the same instrument. This
reduces the number of instruments required to analyze
CSF leading to a reduction in costs.
2.1.5. Hemoglobin and its variants
Analysis of hemoglobinopathies and thalassemias
is important in the diagnosis and management of the
over 600 known congenital hemoglobin (Hb) defects
[72]. Analysis requires identification and quantifica-
tion of the abnormal Hb along with the minor Hb
components [73]. Historically, these variants have
been characterized by electrophoretic separation using
alkaline CAE or AGE combined with citrate agar at
acid pH. However, these techniques are being replaced
by high performance cation exchange chromatography
(HPCEC) [74 – 76] and affinity methods. More
recently, CZE [77 –84] and CIEF [85 –96] have been
found to be helpful in making the differential diagnosis
of a variety of hemoglobinopathies such as S/beta
thalassemia, G-Philadelphia trait, S/C-Harlem disease,
etc. Much of the pioneering work with CIEF identi-
fication of hemoglobinopathies was done by Hempe,
Craver et al. [85,88,89,95]. In this initial work, they
were able to identify the four most common variants
(C, S, D, G), but could not separate Hb E, O-Harlem,
and O-Arab due to the similar isoelectric points (pI).
However, use of family history along with a sickle
solubility test for O-Harlem could differentiate these
variants. Using a standard sample containing known
Hb variants and constructing a linear regression equa-
tion of pI vs. elution time, the pI of unknown variant
hemoglobins can be calculated. The calculated pI is
then used to help identify the unknown Hb. Hempe et
al. [89] also used CIEF to analyze the isoelectric points
of a number of common and uncommon variants using
patient samples previously verified by a reference
laboratory, commercial control samples, and profi-
ciency testing samples. As with most CE methods,
imprecision due to changes in the electroosmotic flow
(EOF) can be a problem. By running a standard sample
each day in addition to using the Hb A present in each
sample as the reference peak, Hempe et al. was able to
correct for changes in the EOF. In a separate set of
experiments, Mohammad et al. added two pI markers,
one at 6.6 and the other at 7.7, to samples before
performing CIEF [91]. Using these markers, a migra-
tion index, which related the migration time of the Hb
variant to the migration times of the two markers, was
calculated. This reduced the migration CVof the Hb C,
S, F, and A by approximately 4-fold from f 16%
to f 4%.
CIEF can also simultaneously detect the concen-
trations of minor Hb variants such as Hb A2 and Hb F,
which are useful not only in the diagnosis of thalasse-
mia syndromes but also in the long-term follow-up of
sickle cell patients being treated with hydroxylurea.
While it is possible to quantify Hb A2 and Hb F using
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
AGE or HPCEC alone, these methods are commonly
supplemented by minicolumn ion exchange chroma-
tography or alkali denaturation. In contrast, CIEF re-
quires no other analytical procedure in the evaluation
of patients with suspected hemoglobinopathies. Since
minimal follow-up testing is required, use of a refer-
ence laboratory is a cost-effective way to identify the
rarer Hb variants or, as necessary, confirm the identi-
fication made by CIEF [95].
CIEF can also be used to assess glycemic control
through the measurement of Hb A1c, a chemically
modified (glycated) form of normal adult Hb A. The
Diabetes Control and Complications Trial [97] has
shown that Hb A1c levels are a good long-term indica-
tor of the average blood glucose and should be rou-
tinely monitored to check that diabetes is being
properly controlled. Since Hb A1c results can vary with
different analytical methods, national and international
programs have been developed for standardization
[98,99]. Although most laboratories use commercial
HPLC or immunoassay to detect Hb A1c, problems
with these assays have been noted [100]. Recently,
CIEF has been used for separation and quantification of
Hb A1c using the same conditions for separation and
analysis of Hb variants. The method was shown to be
highly correlated (r>0.98) with the gold standard
HPLC method used by the Diabetes Control and
Complications Trial [95,101]. A fixed negative bias
of 1.4% however, was noted, indicating that other
hemoglobin derivatives, such as carbamylated hemo-
globins, may be co-eluting with the Hb A1c in the
HPLC system [101]. Similar findings were also seen by
Hempe et al. [95]. In addition, CIEF can also identify
hemoglobin oxidation products formed during im-
proper storage that can impose bias on Hb A1c results.
Commercially available reagents used in CZE to sep-
arate and quantify Hb A1c have also been evaluated
[101]. While the method is rapid ( < 4 min), relatively
precise (CV < 4%), and unaffected by carbamylated
hemoglobins and hemoglobin variants F, C and S, a
separate proprietary buffer system is required to ana-
lyze the major hemoglobin variants. One benefit of
using CIEF for Hb A1c is that the same instrument and
reagents can also be used to separate hemoglobin
variants. With the increased throughput available with
multi-capillary instruments, CIEF should be able to
compete with HPLC, since only one method and in-
strument is required for these analytes.
9
2.1.6. Carbohydrate-deficient transferrin
Transferrin is an iron transport protein known to be
microheterogeneous in carbohydrate and the terminat-
ing sialic residues [102]. The predominant sialioform in
normal individuals is tetrasialotransferrin, although
minor components are also present at lower concen-
trations [103]. The minor sialioforms, specifically
those with two or less sialic acids, also known as
carbohydrate-deficient transferrin or CDT, have been
shown to be increased in chronic abusers of alcohol
[104]. Studies have shown that CDT can detect exces-
sive alcohol consumption (sensitivity of f 80%) when
a person has consumed an average of >50 g alcohol per
day over a time period of 1 week [105]. Later studies,
however, have shown that diagnostic sensitivities of
30– 50% for women and 50 –70% for men are more
realistic [106]. Even though CDT should not be used as
a screening test for chronic alcohol abuse, it may be a
way to monitor abstinence. An excellent review of the
biology and use of CDT and chronic alcohol consump-
tion has recently been published [106].
The reference method for the measurement of CDT
is isoelectric focusing, followed by immunofixation,
and quantification using densitometry [107]. Commer-
cial methods suitable for the routine clinical laboratory
using anion-exchange in conjunction with immuno-
assay are also available [108– 110], although CDT
recovery can be problematic [110]. Commercial meth-
ods combine all CDT sialoforms into one fraction and
then quantify by immunoassay potentially obscuring
information useful to the clinician [111]. CE which can
examine the CDT profile via direct electrophoresis of
serum may offer a solution to these problems [112 –
118]. The need for minimal sample preparation and the
ability to directly detect the various sialoforms of CDT
decreases analysis time compared to isoelectric focus-
ing separations or commercial assays. As with other
CE-based methods, protein – capillary wall interactions
can be a problem but this challenge can be reduced by
using buffer additives, such as 1,4-diaminobutane
[116,117], proprietary buffers [118], or coated capil-
laries [116 – 119]. Coated capillaries work well [112 –
115] but can give unpredictable results. Recently,
Crivellente et al. [116] described a CE method to
separate CDT, using 1,4-diaminobutane to dynami-
cally coat the capillary surface. Although the separa-
tion of CDT was improved over their previous method
[115], they were not able to detect the asialo isoform in
10 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30

a sample containing 61 U/l CDT. This is in contrast to
IEF which can detect asialiotransferrin in samples con-
taining CDT at much lower levels [119]. Giordano et al.
[117] also used 1,4-diaminobutane and fused silica
capillaries to separate the sialoforms of transferrin with
similar results. These researchers were able to obtain a
cleaner and more easily interpreted electropherogram
by using protein A covalently attached to agarose to
remove the IgG, but still were not able to see asialio-
transferrin. It is possible that an even cleaner electro-
pherogram may be obtained using the genetically
engineered hybrid of Protein L and Protein A that
binds all immunoglobulins subclasses including free
kappa light chain [120]. This may allow better visual-
ization of asialiotransferrin. While research methods
were not able to detect asialiotransferrin, use of a
commercial kit (CEofixk CDT kit, Analis, Belgium)
and the Paragon CZE 2000 detected asialiotransferrin
(Fig. 3). Although additional studies are necessary to
validate the use of CE in the detection and quantifica-
tion of CDT, it appears to be an acceptable replacement
for current methods in the evaluation of excessive
alcohol consumption.
2.1.7. Lipoproteins
Atherosclerosis, a chronic disease characterized by
the localized accumulation of plaque, is the principal
cause of coronary arterial disease leading to the ob-
struction of arteries [121]. Serum lipoprotein abnorma-
lities, such as increased LDL, decreased HDL, in-
creased Lp (a), etc., have been shown to be a factor
in atherosclerotic development [122,123]. Because of
this association, lipoprotein profiles consisting of
serum cholesterol, triglycerides, HDL, and LDL (cal-
culated using the Friedewald formula) are routinely
run in the main clinical chemistry laboratory. It is pos-
sible to obtain the same information along with addi-
tional information on Lp (a), IDL, VLDL, LDL, and

Fig. 3. CEofixk CDT kit for determination of carbohydrate deficient transferrin (CDT) using the Beckman Paragon CZER 2000. Separation
conditions: fused silica capillary with a capillary length is 57 cm (50 cm to detector)  50 Am; temperature, 40 jC, a proprietary buffer (CEofix
CDT reagent set) was used at 28 kV for 7 min; injection by 0.5 psi for 1 s. The electrophoregram has been expanded to show the region where
CDT is found. Results are shown for three social drinkers (electrophorograms labelled 1 – 2) and four heavy drinkers (electropherograms labelled
4 – 7). Peaks detected: (0) asialotransferrin, (2) disialotransferrin, (3) trisialotransferrin, (4) tetrasialotransferrin, (5) pentasialotransferrin, (6)
hexasialotransferrin. Asialotransferrin is absent in the social drinkers but can be detected in the heavy drinkers. In addition, disialotransferrin can
be seen to be increased in the heavy drinkers relative to the social drinkers. The figure was kindly supplied by Beckman Coulter.
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
HDL by using AGE along with enzymatic staining of
cholesterol to directly quantify the lipoprotein frac-
tions [124]. Similar to other electrophoretic methods,
CE can replace AGE in the separation and detection of
all serum lipoproteins as discussed below. Using
capillary isotachophoresis (CITP), it is possible to
separate serum lipoproteins into 9 – 14 fractions
[125 –133]. Using Sudan Black as a lipoprotein stain,
Schmitz and Möllers [125] were able to show that
various hyperlipoproteinemias, classified according to
the Frederickson classification, were clearly distin-
guishable by CITP. In addition, they were able to
observe the effect of drug therapy on the various
lipoprotein fractions. Recently, Schmitz et al. [126]
used the lipophilic fluorescent dye, 7-nitrobenz-2-oxa-
1,3-diazole ceramide, to stain lipoproteins. Although
the absolute values were not identical, reasonable
correlations were found for HDL and LDL (r = 0.9
and 0.91, respectively) vs. routinely used methods for
a small number of patients. Zorn et al. [127] used either
an enzymatic specific staining of cholesterol or trigly-
ceride to distinguish the various lipoprotein fractions.
CITP has been shown to separate serum lipoproteins
into multiple fractions, including at least five HDL and
seven LDL subfractions. Once the clinical usefulness
in monitoring these additional subfractions has been
established, CITP should become the method of choice
in monitoring disorders of lipoprotein metabolism. In
addition, CITP can also be used to identify apolipo-
proteins directly from serum [129,134], however, sig-
nificant sample preparation is required limiting it
usefulness in the clinical laboratory.
2.2. Serum and urine analysis of drugs
Studies have shown that CE has better resolution,
reduced sample preparation, costs less, and is faster
than high performance liquid chromatography (HPLC)
[136 –142], although the sensitivity and migration time
precision for CE is not good as HPLC [135]. Sensitivity
can be increased by using extended light pathlength
capillaries, such as bubble cells or z-cells, where the
inner diameter of the capillary is increased only at the
detection window. Although extended light pathlength
capillaries appear to work most researchers rely on
field-amplified stacking where increases in sensitivity
of up to a 1000 increase have been attained [143].
Stacking also improves the precision of analyte quan-
11
titation by increasing the amount of analyte on the
capillary [144]. These and other ways to improve the
sensitivity of CE has been recently reviewed by Hem-
pel [145].
In addition to less-than-desirable sensitivity, repro-
ducibility of analyte migration times in CE can also be
poor, especially when MEKC or CZE are used to
separate analytes. This is mainly due to the inability
to control the EOF and is more pronounced when a
capillary is first used. Although attempts to control
EOF has met with limited success, the use of internal
marker(s) to calculate an effective mobility (leff),
which is independent of the EOF, has made it possible
to account for these changes [146 – 149]. This has
reduced the relative standard deviation (RSD) by over
3-fold from 3 –5% to 0.7 – 1.5%. More recently, a new
term—corrected leff—which uses standards with
known leff, has been introduced [150 –152]. The leff
of each standard is based on an average of multiple
analyses. A standard curve is prepared and used to
correct the leff obtained for analytes in each experi-
ment. Using the corrected leff has reduced the intra-
instrument RSD to < 3%, inter-instrument RSD to
< 4%, and inter-laboratory to f 3%. Thus, by using
the leff or the corrected leff, the RSD for CE should be
able to favorability compete with HPLC.
CE has been shown to be useful in the area of
therapeutic drug monitoring (TDM). In addition to
analyzing a drug and its metabolites, CE can also be
used to determine the bound and free fraction of drugs,
drug isomers, and a drugs physico-chemical properties
[153]. In TDM, CE will have the most utility for those
drugs that do not have an immunoassay or HPLC
method already developed.
CE is also useful in forensics [154,155] to screen
urine for drugs of abuse, such as opiates [156], barbi-
turates [157], benzodiazepines [158], amphetamines
[159], morphine [160], or impurities in heroin [161]. In
the initial screening stages forensic toxicology labora-
tories are concerned only with identifying which drugs
are present, if any, and are not concerned with quanti-
fication. Once detected, confirmation and quantifica-
tion can be performed using a different method. The
ultimate goal for screening samples is no false positive
or false negative results [162]. However, this is not
realistic. What forensic laboratories do to minimize
false positives is to confirm all positive results by a
definitive method, usually mass spectrometry (MS),
12 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30

which eliminates most false positive results. Although
some false positive results are acceptable in the screen-
ing stage, false negatives should be kept to a minimum
since a negative result will probably end further sample
analysis. Currently immunoassay and HPLC are the
methods of choice in forensic toxicology [163 –167].
Immunoassays, however, often lack specificity and, at
times, sensitivity, while HPLC methods usually require
extensive sample pretreatment leading to modest sam-
ple throughput. CE has been used to separate and
identify a wide variety of drugs; including drugs of
abuse in body fluids [136 –142,153 – 161,168,169] (see
Fig. 4 for an example of the separation of the halluci-
nogenic amines, hydromorphone, morphine, and co-
deine). It has also been interfaced to MS [159,160,170],
making it feasible to be used both as a screening and as
a confirmatory method.
It is also possible to use CE for screening post-
mortem fluids for illicit drugs or elevated levels of
legal drugs [155]. Hudson et al. has shown CE to be
useful in analyzing whole blood specimens (whether
fresh, hemolysed, or putrid) minimizing false nega-
tives using a simple liquid – liquid extraction proce-
dure [155,171,172]. In addition, the disappearance of
analytes due to thermal decomposition or irreversible
absorption to columns is rarely, if ever, observed in
CE. Use of a simple UV or diode array detector basic
drugs allowed the detection of basic drugs at concen-
trations of 10 ng/ml (Fig. 5). They also characterized
the relative mobilities of over 500 basic and neutral
drugs of forensic interest [171,173]. Because of the
ability to detect large number of drugs and a reduction
in the screening cost the Royal Canadian Mounted
Police forensic laboratory in Regina, Saskatchewan,

Fig. 4. Detection of the hallucinogenic amines, hydromorphone, morphine, and codeine, in whole blood by capillary electrophoresis. Whole-blood
samples (1 ml) containing 10 ng/ml of hydromorphone (HM), morphine (M), and codeine (C) were extracted with 5 ml 1-chlorobutane plus 0.2 ml
ammonia. The residue is redissolved in methanol (1% HCl), evaporated and dissolved in 30 AL water. Internal standard is 100 ng/ml nalorphine.
Separation conditions: capillary 70 cm (60 cm to detector)  75 Am fused silica capillary containing 0.4% h-CD in 100 mM phosphate pH 2.38, at
21 kV; temperature, 25 jC; injection at 10 kV for 16 s; detection at 200 nm. The standard curve shows the linear response of hydromorphone over 3
logs. The figure was kindly supplied by J. Hudson, Royal Canadian Mounted Police, Regina, Saskatchewan, Canada.
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30 13

Fig. 5. Quality controls for basic drugs using the Beckman P/ACE MDQ. Whole-blood samples (1 ml) were extracted with 5 ml 1-chlorobutane
plus 0.2 ml ammonia. The residue is redissolved in methanol (1% HCl), evaporated and dissolved in 30 Al water. Separation conditions: capillary,
60 cm (50 cm to detector)  75 Am fused silica capillary containing 100 mM phosphate, pH 2.38, at 18 kV; temperature, 25 jC; injection at 10 kV
for 8 – 16 s; detection at 200 nm. (a) Quality control sample in water. (b) Quality control extract, 10 ng of each drug in porcine blood. (c) Quality
control blank with only 50 ng/ml internal standard added. The figure was kindly supplied by J. Hudson, Royal Canadian Mounted Police, Regina,
Saskatchewan, Canada.
14 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
Canada is now using CE as the preferred method to
screen forensic samples for drugs.
2.3. Molecular diagnostics
Molecular pathology/molecular diagnostics is the
newest and most rapidly growing area in laboratory
medicine, in which the detection, characterization and/
or quantification of nucleic acids is used to assist in the
diagnosis and management of disease. Molecular
assays augment classical laboratory medicine by pro-
viding additional information that could not be ob-
tained using standard methodologies. Currently, mole-
cular pathology can be separated into six areas: (1)
hematology/oncology; (2) solid tumors; (3) genetics;
(4) pharmacogenetics; (5) infectious diseases; and (6)
identity testing/forensics. Molecular pathology labora-
tories currently focus on infectious diseases. This focus
probably will continue for the next several years;
however, interest in other tests is increasing.
CE-based molecular assays combine high sensitiv-
ity with high resolution that will allow it to become an
important and integral part of the molecular pathology
laboratory. CE has the following advantages over
classical slab polyacrylamide gel electrophoresis
(PAGE) in the detection of infectious agents, genetic
polymorphisms, or quantifying gene expression [174]:
(1) reduction in labor costs required cast and manually
load slab gels; (2) small sample size capabilities and
the ability to analyze non-ideal tissue samples such as
archival paraffin embedded formalin fixed tissue; and
(3) automated digital imaging capabilities that can use
either peak height or peak area in a semi-quantitative
or quantitative manner.
The versatility of CE-based molecular platforms
results from its ability to resolve single base changes
for sequencing or larger base pair differences (greater
than 5 bp) for fragment analysis applications.
Sequence analysis is the gold standard for molecular
testing. It yields the greatest amount of information
since it identifies the order of each deoxynucleotide
base of a particular target, usually amplified DNA or
cDNA [175]. Following the initial amplification, a pu-
rification step removes residual deoxynucleotides and
primers from the polymerase chain reaction (PCR)
product. The products are then subjected to a second
round of amplification similar to a typical PCR reac-
tion. In this second reaction, however, deoxynucleo-
tides along with limiting amounts of chain terminating
dideoxynucleotides labeled with a different flouro-
phores on each base are present, resulting in a series
of different colored, truncated DNA products due to a
random incorporation of the dideoxynucleotides in
competition with the normal deoxynucleotides. The
single-base resolving capability of CE permits all four
flourochrome products to be separated simultaneously
(Fig. 6A and B) compared to classical sequencing with
PAGE, where each chain terminator has to be loaded in
separate lanes. The result is a multi-colored ladder
where each color represents a different base. The order
of the bases is then analyzed using secondary software
that characterizes the sequence in terms of identity, and
relatedness to prototypical sequences in a database. In
contrast, fragment analysis incorporates one flouro-
chrome-labeled PCR primer in a standard PCR reaction
after which the product(s) are separated and visualized.
Fragment analysis is more rapid and less expensive
because the flourochrome is incorporated in the initial
PCR reaction thus eliminating the need for sequencing
reactions that incorporate the dye terminating bases.
Interpretation of the fragment analysis data is also
easier than sequence analysis because the fragments
are readily identified by the imaging software and the
need for secondary sequence software analysis is not
required. While less expensive and more rapid, frag-
ment analysis does not provide exact, detailed
sequence data. In addition, PCR fragments can result
from erroneous amplification and thus verification
using sequencing techniques or independent probes is
sometimes recommended for quality assurance purpo-
ses. In these cases, sequencing is often used to confirm
the identity of the PCR product.
2.3.1. Infectious diseases
An important aspect of molecular methods is that
they do not require the presence of viable organisms
permitting the identification of bacteria, viruses, and
fungi that are difficult if not impossible to culture.
Identification of infectious agents can also be used for
both diagnostic and therapeutic purposes [176 – 180].
Although the simplest application of molecular tech-
niques is to detect infectious entities in body fluids
which are nearly sterile [181,182], more complex
analysis is also done, such as detailed characterization
of infectious agents via sequence analysis. For
instance, detection of genetic polymorphisms in tuber-
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30 15

Fig. 6. A. Image of processed CE sequencing data. Raw sequence data was collected for BRCA mutation analysis. Note that different colors
represent different base pair terminators. B. Analysis of BRCA gene mutation by comparison of patient and reference sequences. The patient
sequence is on top with the reference sequence overlaid below. Peaks above and below the line denote changes from the reference sequence.
(Data for Panel B was kindly provided by Tom Frank of Myriad Genetic Laboratories, Salt Lake City, UT).

culosis has been shown to be helpful in identifying
rifampicin resistant isolates [178] while determination
of the genotypes for Human Immunodeficiency Virus
(HIV) or Hepatitis C Virus (HCV) can predict longev-
ity and treatment modalities using antiviral therapies
[183 –185]. Also, if >2000 viral copies/ml of HIV are
present in plasma, it is possible to determine the
susceptibility of the virus to antiviral therapy. This is
done by sequencing the reverse transcriptase and
protease genes of the HIV that are isolated employing
RT-PCR and multiple sequencing primers. The
sequencing results are aligned using a software pro-
gram that overlaps sequences generated from forward
and reverse sequencing primers (Fig. 7). This software
identifies variations in the HIV sequence (lines in the
top diagram) that may be associated with various
antiviral drug resistance.
PCR can also be used to target conserved stretches
of DNA, such as ribosomal DNA (rDNA), resulting in
new applications that can identify infectious agents
16 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
Fig. 7. HIV Genotyping (ABI) shows alignment data of seven sequences using the Viroseq software permitting comprehensive data of both
forward and reverse strands of the viral cDNA.
that cannot be cultured. The entire 16S rDNA se-
quence (1541 bp) for bacteria can be generated using
a single pair of PCR primers and then sequenced using
the Microseq Microbial Identification systems CE
(Applied Biosystems, Foster City, CA). Normally,
only 500 bases are required for routine identification,
although sequencing of the entire rDNA is possible.
Using this approach, various bacterial strains or bio-
types have been identified for Streptococcus sp.,
Mycobacterium sp., coryneform bacterial isolates, as
well as other bacterial species [175,186 – 190]. Identi-
fication of fungi using CE is usually done by amplify-
ing and sequencing the large rDNA subunit because of
the multiple gene copies normally present [191].
Although there are 12 domains present in the fungal
rDNA, domain 2 is the largest (200 –500 bp) and is
routinely used for identification. Sequencing of
domain 2 has allowed strain identification of the fungi
Trichophyton rubrum and Schizophyllum communes
associated with diseases of nail tissue and the sinus
cavity, respectively [188,189]. Using this approach,
Kurzai et al. [190] identified Candida dubliniensis as a
new emerging pathogen. This fungi is often incorrectly
identified as Candidia albicans using classical meth-
ods. Sequencing of rDNA by CE by Baele et al. [179]
is also useful in the monitoring nosocomial infections.
Typically, rDNA various isolates are amplified, se-
quenced, and then compared to determine if the strains
are the same as, or in some manner related to, pre-
viously isolated strains.
2.3.2. Hematology/oncology
CE is useful in the analysis of molecular hematol-
ogy/oncology tests such as B- and T-cell clonality
assays [174,192 –198]. Using CE, monoclonal popu-
lations of B cells are detected are detected through
analysis of changes in the immunoglobulin heavy
chain gene (Fig. 8). Because this region is polymor-
phic, a number of different primers (seven upstream
VH region primers and three JH primers obtained from
InVivoscribe LLC, CA) can be used to increase the
ability to identify clonal heavy chain changes. It is also
possible to identify changes in the Kappa chain using
this approach with different primers. T-cell mono-
clonal populations can be detected using PCR, ampli-
fying either the T-cell gamma and/or beta receptor
[195,197]. Generally, the presence of one or two peaks
with intensities greater than 2-fold over background is
considered positive for a gene rearrangement. Prior to
CE, distinguishing a monoclonal peak from a back-
ground of normal rearrangements was very subjective.
Objective data could only be obtained using densi-
tometry. In contrast, CE cannot only identify a peak
but give numerical data using the peak height or area
helping to facilitate interpretation. However, since
several reports have demonstrated pseudo-spikes
resulting from reactive rather than oncogenic pro-
cesses [199,200], it is important to remember that
identification of monoclonality does not always mean
malignancy. Although CE facilitates interpretation of
these difficult cases, it is critical that each clinical
laboratory perform extensive validation to assure that
interpretation of a rearrangement test is accurate.
Detection of gene translocations by CE has been
used to differentiate various types of lymphomas and
leukemias aiding in the decision of appropriate ther-
apy. In gene translocation assays, PCR uses primers
from the two different chromosomes [194]. In this
manner, mantle cell lymphomas have been found to be
associated with the translocation of BCL1 t (11:14),
follicular lymphomas with BCL2 t (14:18), and chro-
nic myleogenous leukemia with Bcr/Abl t (9:22).
The ability of CE to accurately determine the size of
the monoclonal population and/or the translocation
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30 17

Fig. 8. Fragment analysis of gene changes for B- and T-cell monoclonality (Invivoscribe, LLC). Primers for the B- and T-cell monoclonality were
obtained from Invivoscribe and are propritary. The B-cell primers are directed toward the heavy chain and framework III of the immunoglobulin
gene. T-cell primers are directed toward conserved regions of the T cell gamma receptor. Each PCR product was generated using 35 cycles at
95 jC for 1 min, 55 jC for 30 s, 72 jC for 30 s. Blue spikes represent rearrangements corresponding to different size PCR products resulting
from clonal populations. Monoclonal populations are represented by one or two prominent peaks (A and C) while normal clonal populations
are seen as a single bell shaped curve for B-cell (JH) rearrangements using primers directed toward the immunoglobin heavy chain or as a
bimodal curve using primers targeting the T-cell gamma receptor, respectively (B and D). The red spikes represent the standards.
18 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30

makes it useful for monitoring therapy by determining
if treatment has failed or if reoccurrence is due to a new
primary disease. Berg et al. [201] described a case
involving a T-cell lymphoma resulting from allogenic
bone marrow transplantation that clearly demonstrates
the utility of CE in accurately sizing fragments. In this
case, each of two sisters, one the donor and the other
the recipient, developed an identical T-cell lymphoma
as determined by CE, implicating transfer of the
lymphoma cells from the donor to recipient during
transplantation. In addition to accurately sizing PCR
fragments, CE also has the ability to work with very
small quantities of materials making it especially
useful in cases where there is insufficient patient
material or confounding results are found by using
conventional methodologies.
2.3.3. Solid tumors
Solid tumors can result from genetic polymor-
phisms including translocations, single nucleotide po-
lymorphisms (SNP), loss of heterozygosity (LOH),
and microsatellite instability. Translocations can also
arise in various tumor types and are usually detected
using fragment analysis as mentioned above for
hematology/oncology samples. LOH, also known as
allelic loss, is useful in distinguishing between tumor
recurrence vs. de novo cancer formation in addition
to determining prognosis [202,203]. LOH is known
to occur in various cancers, including head and neck
and colon tumors [204 –207] and can be identified
using fragment analysis. These assays detect chro-
mosomal differences between normal and tumor
tissue. Thus, the presence in tumor tissue of normal
tissue in the form of stromal tissue and blood vessels
can be a source of contamination. As a result,
identification of allelic loss requires calculating the
ratio of band intensity of the two alleles in the tumor
vs. normal tissue. A decrease of >30% is the thresh-
old criteria to identify a specific allele loss. As shown
in Fig. 9, CE provides improved resolution, sizing,
and simplified quantification to determine if LOH
has occurred.
SNPs, on the other hand, are detected by sequenc-
ing and primer extension which can be used for
prognostic purposes in solid tumor cases. For exam-
ple, TP53, which is associated with a ‘‘normal’’
genotype, has been shown to predict enhanced patient
survival in squamous cell carcinomas and rectal
cancers [208,209]. Another example involves the
recent success in the treatment of patients with a

Fig. 9. Use of STR PCR to detect LOH. This figure shows how the ratio of alleles can be used to help in distinguishing normal and tumor tissue
using primers targeting the D1S2883 loci which is located near the gene associated with hereditary prostate cancer. In the normal tissue the ratio of
the different alleles is 1.6. This ratio is greatly increased in the tumor with implicates a loss of the larger allele.
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
specific tyrosine kinase inhibitor, STI571, in gastro-
intestinal stromal tumors expressing mutant c-kit
[210,211]. In addition to being used as a solid tumor
test, detection of SNPs can also be usefully in assess-
ing risk in developing cancer. For example, in addition
to being used to help identify the most appropriate
treatment for these cancers, BRCA 1 and 2 variants can
be used to help identify women at risk for developing
breast and ovarian cancer. Genetic counseling is rec-
ommended for cancer patients to assist in understand-
ing the relative risk for developing breast or ovarian
cancer [213]. The presence of the BCRA 1 and 2
variants in a breast tumor can mean that certain
interventions, such as prophylactic contralateral mas-
tectomy, may be useful in preventing a second primary
cancer [212].
2.3.4. Identity testing/paternity testing/HLA testing
Fragment analysis of short tandem repeat (STR)
PCR products such as those used for LOH studies can
also be used to identify specific individuals in a larger
population [214 – 218]. Identity testing has applica-
tions that range from identifying suitable recipients for
organ transplantation, paternity testing, bone marrow
engraftment analysis, forensic testing, and quality
assurance measures for determining the identity of
mislabeled paraffin embedded tissue or foreign tissue
embedded in the same cassette. Fig. 10 demonstrates
the use of these tests for therapeutic follow-up of a
bone marrow engraftment patient. In panel 3 of this
figure, it can be seen from a 50% mixture of donor
Fig. 10. Use of STR PCR to monitor bone marrow engraftment. Panels 1 and 2 represent the donor and the recipient, respectively. Panel 3
demonstrates a 50% chimera in the patient.
19
and recipient DNA that treatment failure due to the
recipient’s cells repopulation of their bone marrow has
probably occurred. Identifying the cause of graft fail-
ure whether due to rejection, graft vs. host, or other
mechanisms, is essential for deciding on the most
appropriate treatment regimen. The figure also dem-
onstrates the ease of identifying multiple alleles
simultaneously using CE. This is the same basic
technique that is used for forensic and paternity cases,
although with forensic tests additional STRs are
usually required to statistically assure the identity of
the subject.
2.3.5. Genetics
CE has simplified the methodology for performing
both simple and complex genetic tests. Numerous
inherited genetic diseases, such as Cystic Fibrosis
(CF), fragile X, mitochondrial heteroplasmy, spino-
cerebellar ataxia, and genetic variants of cytochrome
P450 that are involved with drug metabolism can be
detected using CE [219 – 228]. Sequencing and frag-
ment analysis of restriction fragment length polymor-
phisms (RFLP) and primer extension are commonly
used to detect these variants. Fig. 11 demonstrates the
results of DNA that is heterozygous for a splice
variant of CYP3a5. This splice variant has decreased
activity for some substrates such as protease inhib-
itors, which can potentially result in toxic drug levels.
Using the same technology, it is also possible to detect
the presence of certain genetic mutations that are
associated with an increased risk having a child with
20 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30

Fig. 11. Single nucleotide polymorphism detection using primer extension (Snapshot, ABI) and fragment analysis. Wild-type base pair is red
while the variant is black. (A) CYP3A5*3 Splice Variant 1, heterozygote; (B) CYP3A5*3 Splice Variant 2; (C) Simultaneous detection of
heteroxygote compound for both CYP3A5*3 splice variants.

CF [229]. Currently, the American College of Gyne-
cologists (ACOG) recommends screening for all Cau-
casian women and their partners for the presence of
variants which over have been shown to have a causal
association with CF. For routine screening purposes,
however, screening of only the most common 25
specific variants has been recommended. Although
the use of CE for complex inherited genetic tests has
simplified testing, tremendous amounts of data are
generated [227], necessitating the need for interpreta-
tive guidelines. This has led the College of American
Pathologists (CAP) to recommend that laboratory
reports include the following information to assist in
test interpretation: (1) an estimate of the risk for false
negatives and false positives arising from recombina-
tion between the probe and the disease associated
gene; (2) an estimate of the residual risk of being a
carrier for one of the variants not tested for; and (3)
discussion of recessive or dominant inheritance, recur-
rence risk, penetrance, severity, and other genotype –
phenotype correlatives. In addition to genetic coun-
seling, the laboratory report is necessary to effectively
relay residual risks and uncertainties, and the medical
and reproductive options suggested by the test results
[213].
2.3.6. Other
Currently, gene expression profiles using arrays
permit pattern identification that may be useful for
diagnostic and prognostic purposes. Classically, mem-
brane and silicon arrays identified various patterns but
required specialized equipment to perform and inter-
pret the results. More recently, expression arrays for-
mulated with internal expression controls have enabled
analysis using standard CE equipment [230 –234]. The
advantages of automated quantification, as seen in
previous CE-based clinical tests facilitate its applica-
tion for high throughput analysis of gene expression.
Other applications that hold promise for clinical appli-
cations involve microfluidic chambers [225,235 –
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30 21

237]. As shown in Fig. 12, these chambers can resolve 3. Conclusion

DNA fragments with high resolution. Although these
microfluidic devices could be applied to any point of
care test, in the foreseeable future they will probably
be used either for molecular analysis or identification
of infectious agents.
CE is a sensitive and versatile technique and
represents an inexpensive and practical method for
the determination of many clinically important ana-
lytes. Although CE may be maturing in the research,

Fig. 12. Microfluidic chamber (iChip) layout and an example of the resolution by chip and standard gel electrophoresis. The figure was kindly
provided by Hitachi Chemical (Japan).
22 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
genomic, and pharmaceutical arenas, in the clinical
laboratory, it still appears to be in its early childhood.
Initially, CE was heralded for its speed and low
sample volume capabilities, and perhaps more impor-
tantly, its ability to be quantitative and to automate,
as well as separating compounds that have been
difficult to handle by traditional methods. As recently
stated by one of the major contributors to the area of
CE, Dr. James Jorgenson, CE is the ‘‘best-kept secret
in chemical analysis’’ since it can be used to analyze
complex mixtures and is responsible for virtually all
microfluidics for lab-on-a-chip devices [173]. It is
also possible to use one instrument for multiple
purposes, such as serum, urine, and CSF protein;
hemoglobin (both variant and A1c); lipoprotein;
carbohydrate-deficient transferrin; and alpha-1 anti-
trypsin deficiency analysis. The main advantage of
CE is its ability to screen large numbers of specimens
with very little hands-on time at a relatively low cost
per test.
Although analysis of serum proteins by CE offers
many advantages in terms of labor saving per test
and reproducibility, it has not been generally accep-
ted in the clinical laboratory. This is clearly seen by
looking at the number of laboratories subscribing to
CAP proficiency testing for serum proteins electro-
phoresis. In 200,1 the number of laboratories using
CE for serum proteins was 13 out of a total of 739.
So why is CE not being used in the routine clinical
laboratory? Possibly because CE is still perceived as
requiring above-average technical expertise preclud-
ing its use in a laboratory workforce that is less
technically adept.
Broader acceptance of CE in the main clinical
laboratory will require the following: (1) the devel-
opment of chemical reagent kits, instruments and
software suitable for daily clinical laboratory use;
(2) an increase of sensitivity and sample throughput;
and (3) comparison and evaluation of various appli-
cations with current laboratory methods under daily
laboratory conditions. Thus, a greater role for CE in
the future clinical laboratory can be anticipated as
the ease of use, number, variety of applications
increase.
Although CE has not been accepted in the main
clinical laboratory, the technique is being used in the
molecular pathology laboratory. Its use, however, is
not apparent by simply looking at CAP proficiency
surveys, which indicate that only 12 of 70 molecular
pathology laboratories use CE. From our experience,
this did not appear to be correct. Thus, we initiated
an informal survey of 33 molecular pathology labo-
ratories, which showed that all but three use CE in
some part of their testing. CE has simplified molec-
ular tests by providing a platform that can address all
areas of molecular testing. Because of its ease of use
and robust nature, it is becoming a standard piece of
equipment for every molecular pathology laboratory.
In the future, CE will be the basis of additional
molecular techniques, especially in the area of
expression arrays.
References
[1] Lehmann R, Voelter W, Liebich HM. Capillary electrophore-
sis in clinical chemistry. J Chromatogr B 1997;697:3 – 35.
[2] Shihabi ZK, Friedberg MA. Analysis of small molecules for
clinical diagnosis by capillary electrophoresis. Electrophore-
sis 1997;18:1724 – 32.
[3] von Heeren F, Thormann W. Capillary electrophoresis in
clinical and forensic analysis. Electrophoresis 1997;18:
2415 – 26.
[4] Thormann W, Aebi Y, Lanz M, Caslavska J. Capillary electro-
phoresis in clinical toxicology. Forensic Sci Int 1998;92:
157 – 83.
[5] Perrett D. Capillary electrophoresis in clinical chemistry. Ann
Clin Biochem 1999;36:133 – 50.
[6] Altria KD. Overview of capillary electrophoresis and ca-
pillary electrochromatography. J Chromatogr A 1999;856:
443 – 63.
[7] Maurer HH. Systematic toxicological analysis procedures for
acidic drugs and/or metabolites relevant to clinical and foren-
sic toxicology and/or doping control. J Chromatogr B 1999;
733:3 – 25.
[8] Thormann W, Wey AB, Lurie IS, Gerber H, Byland C, Malik
N, et al. Capillary electrophoresis in clinical and forensic
analysis: recent advances and breakthrough to routine appli-
cations. Electrophoresis 1999;20:3203 – 36.
[9] Blessum C, Jeppsson JO, Aguzzi F, Bernon H, Bienvenu J.
Capillary electrophoresis: principles and practice in the clin-
ical laboratory. Ann Biol Clin 1999;57:643 – 57.
[10] Issaq HJ. A decade of capillary electrophoresis. Electropho-
resis 2000;21:1921 – 39.
[11] Jenkins MA. Clinical applications of capillary electrophore-
sis. Status at the new millennium. Mol Biotechnol 2000;15:
201 – 9.
[12] Jin LL, Ferrance J, Landers JP. Miniaturized electrophoresis:
an evolving role in laboratory medicine. BioTech 2001;31:
1332 – 53.
[13] Palfrey SM, editor. Clinical applications of capillary electro-
phoresis. Totowa, NJ: Humana Press; 1999.
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
[14] Petersen JR, Mohammad AA, editors. Clinical and forensic
applications of capillary electrophoresis. Totowa, NJ: Hu-
mana Press; 2001.
[15] Jorgenson JW, Lukacs KD. Zone electrophoresis in open
tubular glass capillaries. Anal Chem 1981;53:1298 – 302.
[16] Kyle RA, Therneau TM, Rajkumar SV, Offord JR, Larson
DR, Plevak MF, et al. Long-term study of prognosis in mono-
clonal gammopathy of undetermined significance. N Engl J
Med 2002;346(8):564 – 9.
[17] Hjerten S. Zone broadening in electrophoresis with special
reference to high-performance electrophoresis in capillaries:
an interplay between theory and practice. Electrophoresis
1990;11(9):665 – 90.
[18] Hjerten S, Kubo K. A new type of pH- and detergent-stable
coating for elimination of electroendosmosis and adsorption in
(capillary) electrophoresis. Electrophoresis 1993;14(5 – 6):
390 – 5.
[19] Gordon MJ, Lee KJ, Arias AA, Zare RN. Protocol for re-
solving protein mixtures in capillary zone electrophoresis.
Anal Chem 1991;63(1):69 – 72.
[20] Chen FT, Liu CM, Hsieh YZ, Sternberg JC. Capillary electro-
phoresis—a new clinical tool. Clin Chem 1991;37(1):14 – 9.
[21] Jenkins MA, Guerin MD. Quantification of serum proteins
using capillary electrophoresis. Ann Clin Biochem 1995;32:
493 – 7.
[22] Jenkins MA, Kulinskaya E, Martin HD, Guerin MD. Evalua-
tion of serum protein separation by capillary electrophoresis:
prospective analysis of 1000 specimens. J Chromatogr B
1995;672:241 – 51.
[23] Jenkins MA, Guerin MD. Optimization of serum protein
separation by capillary electrophoresis. Clin Chem 1996;
42:886.
[24] Jenkins MA, Guerin MD. Capillary electrophoresis proce-
dures for serum protein analysis: comparison with estab-
lished techniques. J Chromatogr B 1997;699:257 – 68.
[25] Katzmann JA, Clark R, Sanders E, Landers JP, Kyle RA.
Prospective study of serum protein capillary zone electro-
phoresis and immunotyping of monoclonal proteins by im-
munosubtraction. Am J Clin Pathol 1998;110:503 – 9.
[26] Bossuyt X, Bogaerts A, Schiettekatte G, Blanckaert N. De-
tection and classification of paraproteins by capillary immu-
nofixation/subtraction. Clin Chem 1998;44:760 – 4.
[27] Bossuyt X, Mariën G. False-negative results in detection of
monoclonal proteins by capillary zone electrophoresis: a pro-
spective study. Clin Chem 2001;47:1477 – 9.
[28] Jonsson M, Carlson J, Jeppsson JO, Simonsson P. Computer-
supported detection of M-components and evaluation of im-
munoglobulins after capillary electrophoresis. Clin Chem
2001;47(1):110 – 7.
[29] Jolliff CR, Blessum C. Comparison of serum protein electro-
phoresis by agarose gel and capillary zone electrophoresis in
a clinical setting. Electrophoresis 1997;18:1781 – 4.
[30] Bossuyt X, Schiettekatte G, Bogaerts A, Blanckaert N. Se-
rum protein electrophoresis by CZE 2000 clinical capillary
electrophoresis system. Clin Chem 1998;44:749 – 59.
[31] Petrini C, Alessio MG, Scapellato L, Brambilla S, Franzini C.
Serum proteins by capillary zone electrophoresis: approaches
23
to the definition of reference values. Clin Chem Lab Med
1999;37:975 – 80.
[32] Winen PAHM, van Dieijen-Vesser MP. Capillary electropho-
resis of serum protein. Reproducibility, comparison with
agarose gel electrophoresis and a review of the literature.
Eur J Clin Chem Clin Biochem 1996;34:535 – 45.
[33] Wong MS, Aw TC. A preliminary evaluation of serum pro-
teins by capillary electrophoresis. J Capillary Electrophor
1998;5:217 – 21.
[34] Clark R, Katzmann JA, Wiegert E, Namyst-Goldberg C,
Sanders L, Oda RP, et al. Rapid capillary electrophoretic
analysis of human serum proteins: qualitative comparison
with high-throughput agarose gel electrophoresis. J Chroma-
togr A 1996;744:205 – 13.
[35] Katzmann JA, Clark R, Wiegert E, Sanders E, Oda RP, Kyle
RA, et al. Identification of monoclonal proteins in serum: a
quantitative comparison of acetate, agarose gel, and capillary
electrophoresis. Electrophoresis 1997;l8:1775 – 80.
[36] Godey F, Ropert M, Bouasria A, Lucas-Clerc C, Guenet L,
Savoure N, et al. Capillary electrophoresis with the Paragon
CZE2000: evaluation and comparison with the gel electro-
phoresis system Hydrasys for serum protein electrophoresis
and monoclonal component typing. Ann Biol Clin 1999;57:
337 – 44.
[37] Doelman CJ, Siebelder CW, Penders TJ. Routine serum pro-
tein analysis using capillary electrophoresis. Eur J Clin Chem
Clin Biochem 1997;35:393 – 4.
[38] Mirura T, Funato T, Yabuki S, Sasaki TS, Kaku MT. Detec-
tion of monoclonal proteins by capillary electrophoresis using
a Zwitterion in the running buffer. Clin Chim Acta 2000;
299:87 – 99.
[39] Bossuyt X, Claeys R, Bogaert G, Said HI, Wouters C, Groven
C, et al. Reference values for the five electrophoretic serum
protein fractions in Caucasian children by capillary zone elec-
trophoresis. Clin Chem Lab Med 2001;39:970 – 2.
[40] Gonzalez-Sagrado M, Lopez-Hernandez S, Martin-Gil FJ,
Tasende J, Banuelos MC, Fernandez-Garcia N, et al. Al-
pha1-antitrypsin deficiencies masked by a clinical capillary
electrophoresis system (CZE 2000). Clin Biochem 2000;33:
79 – 80.
[41] Lupi A, Viglio S, Luisetti M, Gorrini M, Coni P, Faa G, et al.
a1-Antitrypsin in serum determined by capillary isoelectric
focusing. Electrophoresis 2000;21:3318 – 26.
[42] Duc J, Morel B, Peitrequin RB, Frei PC. Identification of
monoclonal gammopathies: a comparison of immunofixation,
immunoelectrophoresis and measurements of n- and E- im-
munoglobulin levels. J Clin Lab Immunol 1988;26:141 – 6.
[43] Guinan JEC, Kenny DF, Gatenby P. Detection and typing of
paraproteins: comparison of different methods in a routine
diagnostic laboratory. Path 1989;21:35 – 41.
[44] Chu SY, MacLeod JE, Bocci L, Monteith M. Characterization
of small monoclonal protein bands with Beckman’s ‘‘Para-
gon’’ immunofixation system. Clin Chem 1987;33:617.
[45] Aguzzi F, Poggi N. Immunosubstraction electrophoresis: a
simple method for identifying specific proteins producing
the cellulose acetate electropherogram. Bull Inst Sierater Mi-
lan 1977;29:7 – 19.
24 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
[46] Klein GL, Joliff CR. Capillary electrophoresis for the rou-
tine clinical laboratory. In: Landers JP, editor. Handbook of
capillary electrophoresis. Boca Raton, FL: CRC Press; 1994.
p. 419 – 58.
[47] Clark R, Katzmann JA, Kyle RA, Fleisher M, Landers JP.
Differential diagnosis of gammopathies by capillary electro-
phoresis and immunosubstraction: analysis of serum samples
problematic by agarose gel electrophoresis. Electrophoresis
1998;19:2479 – 84.
[48] Bossuyt X, Bogaerts A, Schiettekatte G, Blanckaert N. De-
tection and classification of paraproteins by capillary immu-
nofixation/subtraction. Clin Chem 1998;44:760 – 4.
[49] Bienvenu J, Graziani MS, Arpin F, Bernon H, Blessum C,
Marchetti C, et al. Multicenter evaluation of the Paragon
CZEk 2000 capillary zone electrophoresis system for serum
protein electrophoresis and monoclonal component typing.
Clin Chem 1998;44:599 – 605.
[50] Henskens Y, de Winter J, Pekelharing M, Ponjee G. Detec-
tion and identification of monoclonal gammopathies by
capillary electrophoresis. Clin Chem 1998;44:1184 – 90.
[51] Litwin CM, Anderson SK, Philipps G, Martins TB, Jaskow-
ski TD, Hill HR. Comparison of capillary zone and immu-
nosubtraction with agarose gel and immunofixation
electrophoresis for detecting and identifying monoclonal
gammopathies. Am J Clin Pathol 1999;112:411 – 7.
[52] Jenkins MA, Ratnaike S. Five unusual serum protein presen-
tations found by capillary electrophoresis in the clinical lab-
oratory. J Biochem Biophys Methods 1999;41:31 – 47.
[53] Shihabi ZK. Analysis and general classification of serum
cryoglobulins by capillary zone electrophoresis. Electropho-
resis 1996;17:1607 – 12.
[54] Jenkins MA. Three methods of capillary electrophoresis com-
pared with high-resolution agarose gel electrophoresis for
serum protein electrophoresis. J Chromatog B 1998;720:
49 – 58.
[55] Claeys R, Croven C, Gorus FK. Capillary zone electropho-
resis of proteins in body fluids: comparison of capillary and
agarose gel electrophoresis. Clin Chem 2001;47:967 – 70.
[56] Light RW, McGregor MI, Luchisinger PC, Ball WC. Pleural
effusions: the diagnostic separation of transudates and exu-
dates. Ann Intern Med 1972;77:507 – 13.
[57] Waller KV, Ward KM, Mahan JD, Wismatt DK. Current
concepts in proteinuria. Clin Chem 1989;35:755 – 65.
[58] Johnson AM, Rohlfs EM, Silverman LW. Proteins. In: Burtis
CA, Ashwood ER, editors. Tietz fundamentals of clinical
chemistry. Philadelphia: Saunders; 2001. p. 325 – 51.
[59] Friedberg MA, Shahabi ZK. Urine protein analysis by capil-
lary electrophoresis. Electrophoresis 1997;18:1836 – 41.
[60] Jenkins MA. Clinical application of capillary electrophoresis
to unconcentrated human urine proteins. Electrophoresis
1997;18:1842 – 6.
[61] Kolios G, Bairaktari E, Tsolas O, Seferiadis K. Routine differ-
ential diagnosis of proteinurias by capillary electrophoresis.
Clin Chem Lab Med 2001;39:784 – 8.
[62] McFarlin DE, McFarlin HF. Multiple sclerosis. N Engl J Med
1982;307:1974 – 7.
[63] Johnson KP, Arrigo SC, Nelson BJ. Agarose electrophoresis
of cerebrospinal in multiple sclerosis. Neurology 1977;l27:
273 – 7.
[64] Link H, Miller R. Immunoglobins in multiple sclerosis and
infections of the nervous system. Arch Neurol 1971;25:
326 – 44.
[65] Olsson JE, Pettersson B. A comparison between agar gel
electrophoresis and CSF serum quotients of IgG and albu-
min in neurological diseases. Acta Neurol Scand 1976;53:
308 – 22.
[66] Cowdrey G, Firth M, Firth G. Separation of cerebrospinal
fluid proteins using capillary electrophoresis: a potential
method for the diagnosis of neurological disorders. Electro-
phoresis 1995;16:1922 – 6.
[67] Hiraoka A, Arato T, Tominaga I, Eguchi N, Oda H, Urade Y.
Analysis of low-molecular-mass proteins in cerebrospinal
fluid by sodium dodecyl sulfate capillary gel electrophoresis.
J Chromatogr B 1997;697:141 – 7.
[68] Hiraoka A, Tominaga I, Hori K. Sodium dodecylsulfate capil-
lary gel electrophoretic measurement of the concentration
ratios of albumin and a2-macroglobulin in cerebrospinal fluid
and serum of patients with neurological disorders. J Chroma-
togr A 2000;895:339 – 44.
[69] Manabe T, Miyamoto H, Inoue K, Nakatsu M, Aral M. Sep-
aration of human cerebrospinal fluid proteins by capillary
isoelectric focusing in the absence of denaturing agents. Elec-
trophoresis 1999;20:3677 – 83.
[70] Sanders E, Katzmann JA, Clark R, Oda RP, Shihabi Z, Land-
ers JP. Development of capillary electrophoresis as an alter-
native to high resolution agarose electrophoresis for the
diagnosis of multiple sclerosis. Clin Chem Lab Med 1998;
37:37 – 45.
[71] Smith GP, Kjeldsberg CR. Cerebrospinal, synovial, and se-
rous body fluids. In: Henry JB, editor. Clinical diagnosis and
management by laboratory methods. Philadelphia: Saunders;
2001. p. 408.
[72] International Hemoglobin Information Center variant list. He-
moglobin 1995;19:37 – 149.
[73] Elghetany MT, Davey FR. Erythrocytic disorders. In: Henry
JB, editor. Clinical diagnosis and management by laboratory
methods. Philadelphia: Saunders; 2001. p. 560 – 71.
[74] Ou CN, Rognerud CL. Rapid analysis of hemoglobin variants
by cation-exchange HPLC. Clin Chem 1993;39:820 – 4.
[75] Riou J, Godart C, Hurtrel D, Mathis M, Bimet C, Barakad-
jian-Michau J, et al. Cation-exchange HPLC evaluated for
presumptive identification of hemoglobin variants. Clin
Chem 1997;43:34 – 9.
[76] Fucharoen S, Winichagoon P, Wisedpanichkij R, Sae-Ngow
B, Sriphanich R, Oncoung W, et al. Prenatal and postnatal
diagnoses of thalassemias and hemoglobinopathies by HPLC.
Clin Chem 1998;44:740 – 8.
[77] Zhu M, Rodriguez R, Wehr T, Siebert C. Capillary electro-
phoresis of hemoglobins and globin chains. J Chromatogr
1992;608:225 – 37.
[78] Ishioka N, Iyori N, Kurioka S. Detection of abnormal haemo-
globin by capillary electrophoresis and structural identifica-
tion. Biomed Chromatogr 1992;6:224 – 6.
[79] Sahin A, Laleli YR, Ortancil R. Haemoglobin analysis by
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
capillary zone electrophoresis. J Chromatogr 1995;709:
121 – 5.
[80] Jenkins MA, Hendy J, Smith IL. Evaluation of hemoglobin
A2 quantification assay and hemoglobin variant screening by
capillary electrophoresis. J Capillary Electrophor 1997;4:
137 – 43.
[81] Mario N, Baudin B, Bruneel A, Janssens J, Vaubourdolle M.
Capillary zone electrophoresis for the diagnosis of congenital
hemoglobinopathies. Clin Chem 1999;45:285 – 8.
[82] Cotton F, Lin C, Fontaine B, Gulbis B, Janssens J, Vertongen
F. Evaluation of a capillary electrophoresis method for rou-
tine determination of hemoglobins A2 and F. Clin Chem
1999;45:237 – 43.
[83] Mario N, Baudin B, Bruneel A, Janssens J, Vaubourdolle M.
Capillary zone electrophoresis for the diagnosis of congenital
hemoglobinopathies. Clin Chem 1999;45:285 – 8.
[84] Gerritsma J, Sinnige D, Drieze C, Sittrop B, Houtsma P,
Hulshorst-Jansen N, et al. Quantitative and qualitative anal-
ysis of haemoglobin variants using capillary zone electro-
phoresis. Ann Clin Biochem 2000;37:380 – 9.
[85] Hempe JM, Craver RD. Quantification of hemoglobin var-
iants by capillary isoelectric focusing. Clin Chem 1994;40:
2288 – 95.
[86] Conti M, Gelfi C, Righetti PG. Screening of umbilical cord
blood hemoglobins by isoelectric focusing in capillaries.
Electrophoresis 1995;16:1485 – 91.
[87] Wu J, Pawliszyn J. Application of capillary isoelectric focus-
ing with absorption imaging detection to the quantitative de-
termination of human hemoglobin variants. Electrophoresis
1995;16:670 – 3.
[88] Craver RD, Abermanis JG, Warrier RP, Ode DL, Hempe JM.
Hemoglobin A2 levels in healthy persons, sickle cell disease,
sickle cell trait, and beta-thalassemia by capillary isoelectric
focusing. Am J Clin Pathol 1997;107:88 – 91.
[89] Hempe JM, Granger JG, Craver RD. Capillary isoelectric
focusing of hemoglobin variants in the pediatric clinical lab-
oratory. Electrophoresis 1997;18:1785 – 95.
[90] Mario N, Baudin B, Aussel C, Giboudeau J. Comparison
between capillary isoelectric focusing and high-performance
cation-exchange chromatography for qualitative and quanti-
tative analysis of hemoglobin variants. Clin Chem 1997;43:
2137 – 42.
[91] Mohammad AA, Okorodudu AO, Bissell MG, Dow P, Reger
G, Meier A, et al. Clinical application of capillary isoelectric
focusing on fused silica capillaries for determination of he-
moglobin variants. Clin Chem 1997;43:1798 – 9.
[92] Jenkins MA, Ratnaike S. Capillary isoelectric focusing of
haemoglobin variants in the clinical laboratory. Clin Chim
Acta 1999;289:121 – 32.
[93] Saccomani A, Gelfi C, Wajcman H, Righetti PG. Detection
of neutral and charged mutations in alpha- and beta-human
globin chains by capillary zone electrophoresis in isoelectric,
acidic buffers. J Chromatog 1999;832:225 – 38.
[94] Sugano M, Hidaka H, Yamauchi K, Nakabayashi T, Higuchi
Y, Fujita K, et al. Analysis of hemoglobin and globin chain
variants by a commonly used capillary isoelectric focusing
method. Electrophoresis 2000;21:3016 – 9.
25
[95] Hempe JM, Vargas A, Craver RD. Clinical analysis of struc-
tural hemoglobin variants and Hb A1c by capillary isoelec-
tric focusing. In: Petersen JR, Mohammad AA, editors.
Clinical and forensic applications of capillary electrophore-
sis. Totowa, NJ: Humana Press; 2001. p. 145 – 63.
[96] Hempe JM, Craver RD. Separation of hemoglobin variants
with similar charge by capillary isoelectric focusing: value of
isoelectric point for identification of common and uncom-
mon hemoglobin variants. Electrophoresis 2000;21:743 – 8.
[97] The Diabetes Control and Complications Trial Research
Group. Diabetes Control and Complications Trial (DCCT):
results of feasibility study. The DCCT research group. Dia-
betes Care 1987;10:1 – 19.
[98] Weykamp CW, Penders TJ, Miedema K, Muskiet FA, van
der Slik W. Standardization of glycohemoglobin results and
reference values in whole blood studied in 103 laboratories
using 20 methods. Clin Chem 1995;41:82 – 6.
[99] Little RR, Wiedmeyer HM, England JD, Wilke AL, Rohlfing
CL, Wians Jr FH, et al. Interlaboratory standardization of
measurements of glycohemoglobins. Clin Chem 1992;38:
2472 – 8.
[100] Goldstein DE, Little RR, Lorenz RA, Malone JI, Nathan D,
Peterson CM. Tests of glycemia in diabetes. Diabetes Care
1995;18:896 – 909.
[101] Doelman CJA, Siebelder CWM, Nijhof WA, Weykamp CW,
Janssens J, Penders TJ. Capillary electrophoresis system for
hemoglobin A1c determinations evaluated. Clin Chem
1997;43:644 – 8.
[102] de Jong G, van Dijk JP, van Eijk HG. The biology of trans-
ferrin. Clin Chim Acta 1990;190:1 – 46.
[103] van Eijk HG, van Noort WL, de Jong G, Koster JF. Human
serum sialo transferrins in diseases. Clin Chim Acta 1987;
165:141 – 5.
[104] Mårtensson O, Härlin A, Brandt R, Seppä K, Sillanaukee P.
Transferrin isoform distribution: gender and alcohol con-
sumption. Alcohol Clin Exp Res 1997;21:1710 – 5.
[105] Stibler H. Carbohydrate-deficient transferrin in serum: a new
marker of potentially harmful alcohol consumption reviewed.
Clin Chem 1991;37:2029 – 37.
[106] Arndt T. Carbohydrate-deficient transferrin as a marker of
chronic alcohol abuse: a critical review of preanalysis, anal-
ysis, and interpretation. Clin Chem 2001;47:13 – 27.
[107] Hackler R, Arndt T, Kleine TO, Gressner AM. Effect of sep-
aration conditions on automated isoelectric focusing of carbo-
hydrate-deficient transferrin and other human isotransferrins
using the PhastSystem. Anal Biochem 1995;230:281 – 9.
[108] Arndt T, Kropf J, Brandt R, Gressner AM, Hackler R, Herold
M. CDTect-RIA and CDTect-EIA for determination of serum
carbohydrate-deficient transferrin compared. Alcohol 1998;
33:639 – 45.
[109] Bean P, Liegmann K, Løvli T, Westby C, Sundrehagen E.
Semiautomated procedures for evaluation of carbohydrate-
deficient transferrin in the diagnosis of alcohol abuse. Clin
Chem 1997;43:983 – 9.
[110] Arndt T, Hackler R, Kleine TO, Gressner AM. Validation by
isoelectric focusing of the anion-exchange isotransferrin frac-
tionation step involved in determination of carbohydrate-de-
26 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
ficient transferrin by the CDTect assay. Clin Chem 1998;44:
27 – 34.
[111] Heinemann A, Sterneck M, Kuhlencordt R, Rogiers X,
Schulz KH, Queen B, et al. Carbohydrate-deficient transfer-
rin: diagnostic efficiency among patients with end-stage liver
disease before and after liver transplantation. Alcohol Clin
Exp Res 1998;22:1806 – 12.
[112] Oda PR, Prasad R, Stout RL, Coffin D, Patton WP, Kraft DL,
et al. Capillary electrophoresis-based separation of transferrin
sialoforms in patients with carbohydrate-deficient glycopro-
tein syndrome. Electrophoresis 1997;18:1819 – 26.
[113] Trout AL, Prasad R, Coffin D, di Martini A, Lane T, Blessum
C, et al. Direct capillary electrophoretic detection of carbo-
hydrate-deficient transferrin in neat serum. Electrophoresis
2000;21:2376 – 83.
[114] Prasad R, Stout RL, Coffin D, Smith J. Analysis of carbohy-
drate deficient transferrin by capillary zone electrophoresis.
Electrophoresis 1997;18:1814 – 8.
[115] Tagliaro F, Crivellente F, Manetto G, Puppi I, Deyl Z, Marigo
M. Optimized determination of carbohydrate-deficient trans-
ferrin isoforms in serum by capillary zone electrophoresis.
Electrophoresis 1998;19:3033 – 9.
[116] Crivellente F, Fracasso G, Valentini R, Manetto G, Riviera
AP, Tagliaro F. Improved method for carbohydrate-deficient
transferrin determination in human serum by capillary zone
electrophoresis. J Chromatogr B 2000;739:81 – 93.
[117] Giordano BC, Muza M, Trout A, Landers JP. Dynamically-
coated capillaries allow for capillary electrophoretic resolu-
tion of transferrin sialoforms via direct analysis of human
serum. J Chromatogr B 2000;742:79 – 89.
[118] Wuyts B, Delanghe JR, Kasvosve I, Wauters A, Neels H,
Janssens J. Determination of carbohydrate-deficient transfer-
rin using capillary zone electrophoresis. Clin Chem 2001;
47:247 – 55.
[119] Arndt T, Hackler R, Müller T, Kleine TO, Gressner AM.
Increased serum concentration of carbohydrate-deficient
transferring in patients with combined pancreases and kidney
transplantation. Clin Chem 1997;43:344 – 51.
[120] Svensson HG, Hoogenboom HR, Sjobring U. Protein LA, a
novel hybrid protein with unique single-chain Fv antibody-
and Fab-binding properties. Eur J Biochem 1998;258:890 – 6.
[121] McGorisk GM, Treasure CB. Endothelial dysfunction in cor-
onary heart disease. Curr Opin Cardiol 1996;11:341 – 50.
[122] Avins AL, Browner WS. Improving the prediction of coro-
nary heart disease to aid in the management of high choles-
terol levels: what a difference a decade makes. JAMA J Am
Med Assoc 1998;279:445 – 9.
[123] McNamara JR, Shah PK, Nakajima K, Cupples LA, Wilson
PW, Ordovas JM, et al. Remnant lipoprotein cholesterol and
triglyceride reference ranges from the Framingham Heart
Study. Clin Chem 1998;44:1224 – 32.
[124] Nauck M, Winkler K, Marz W, Wieland H. Quantitative
determination of high-, low-, and very-low-density lipopro-
teins and lipoprotein(a) by agarose gel electrophoresis and
enzymatic cholesterol staining. Clin Chem 1995;41:1761 – 7.
[125] Schmitz G, Möllers C. Analysis of lipoproteins with analytical
capillary isotachophoresis. Electrophoresis 1994;15:31 – 9.
[126] Schmitz G, Mollers C, Richter V. Analytical capillary isota-
chophoresis of human serum lipoproteins. Electrophoresis
1997;18:1807 – 13.
[127] Zorn U, Wolf CF, Wennauer R, Bachem MG, Grunert A.
Separation of lipoproteins by capillary isotachophoresis com-
bined with enzymatic derivatization of cholesterol and trigly-
cerides. Electrophoresis 1999;20:1619 – 26.
[128] Bittolo-Bon G, Cazzolato G. Analytical capillary isotacho-
phoresis of total plasma lipoproteins: a new tool to identify
atherogenic low density lipoproteins. J Lipid Res 1999;40:
170 – 7.
[129] Stocks J, Miller NE. Analysis of apolipoproteins and lipopro-
teins by capillary electrophoresis. Electrophoresis 1999;20:
2118 – 23.
[130] Stocks J, Miller NE. Capillary electrophoresis to monitor the
oxidative modification of low densitivity lipoproteins. J Lipid
Res 1998;39:1305 – 9.
[131] Schlenck A, Herbeth B, Siest G, Visvikis S. Characteriza-
tion and quantification of serum lipoprotein subfractions by
capillary isotachophoresis: relationships with lipid, apolipo-
protein, and lipoprotein levels. J Lipid Res 1999;40:
2125 – 33.
[132] Inano K, Tezuka S, Miida T, Okada M. Capillary isotacho-
phoretic analysis of serum lipoproteins using a carrier am-
pholyte as spacer ion. Ann Clin Biochem 2000;37:708 – 16.
[133] Haug ZU, Celik C, Wennauer E, Schmid-Kotsas R, Bachem
A, Grunert MG. Characterization of modified low density
lipoprotein subfractions by capillary isotachophoresis. Elec-
trophoresis 2001;22:1143 – 9.
[134] Lehmann R. Lipoprotein analysis. In: Petersen JR, Mo-
hammad AA, editors. Clinical and forensic applications of
capillary electrophoresis. Totowa, NJ: Humana Press; 2001.
p. 113 – 44.
[135] Wynia GS, Windhorst PC, Post PC, Maris FA. Development
and validation of a capillary electrophoresis method within
pharmaceutical quality control environment and comparison
with high -performance liquid chromatography. J Chroma-
togr A 1997;773:339 – 50.
[136] Shihabi ZK, Oles KS. Felbamate measured in serum by two
methods: HPLC and capillary electrophoresis method. Clin
Chem 1994;40:1904 – 8.
[137] Friedberg MA, Shihabi ZK. Ketoprofen analysis in serum by
capillary electrophoresis. J Chromatogr B 1997;695:193 – 8.
[138] Scholl JB, DeZwaan J. Micellar electrokinetic chromatogra-
phy as a generalized alternative to high-performance liquid
chromatography for purity determination of a class of inves-
tigational antibacterial drugs. J Chromatogr B 1997;695:
147 – 56.
[139] Hsu LC, Constable DJ, Orvos DR, Hannah RE. Comparison
of high-performance liquid chromatography and capillary
zone electrophoresis in penciclovir biodegradation kinetic
studies. J Chromatogr B 1995;669:85 – 92.
[140] Tomlinson AJ, Benson LM, Landers JP, Scanlan GF, Fang J,
Gorrod JW, et al. Investigation of metabolism of the neuro-
leptic drug haloperidol by capillary electrophoresis. J Chro-
matogr A 1993;652:417 – 26.
[141] Heeren F, Tanner R, Theurillat R, Thormann W. Determina-
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
tion of Fluconazole in human plasma by micellar electro-
kinetic capillary chromatography with detection at 190. J
Chromatogr A 1996;745:165 – 72.
[142] Altria KD, Bestford J. Main component assay of pharma-
ceuticals by capillary electrophoresis: considerations regard-
ing precision, accuracy, and linearity data. J Capillary
Electrophor 1996;3:13 – 23.
[143] Zhang C, Thormann W. Head-column field-amplified sample
stacking in binary system capillary electrophoresis: a robust
approach providing over 1000-fold sensitivity enhancement.
Anal Chem 1996;68:2523 – 32.
[144] Shihabi Z. Effect of sample matrix on capillary electro-
phoresis. In: Landers JP, editor. Handbook of capillary
electrophoresis, 2nd ed. Boca Raton, FL: CRC Press;
1997. p. 457 – 77.
[145] Hempel G. Strategies to improve the sensitivity in capillary
electrophoresis for the analysis of drugs in biological fluids.
Electrophoresis 2000;21:691 – 8.
[146] Jumppanen JH, Riekkola M-L. Marker techniques for high-
accuracy identification in CZE. Anal Chem 1995;67:
1060 – 6.
[147] Williams BA, Vigh G. Fast, accurate mobility determination
method for capillary electrophoresis. Anal Chem 1966;68:
1174 – 80.
[148] Yang J, Bose S, Hage DS. Improved reproducibility in capil-
lary electrophoresis through the use of mobility and migra-
tion time ratios. J Chromatogr A 1996;735:209 – 20.
[149] Schmitt-Kopplin P, Garmash AV, Kudryavtsev AV, Men-
zinger F, Perminova IV, Hertkorn N, et al. Quantitative and
qualitative precision improvements by effective mobility-
scale data transformation in capillary electrophoresis analy-
sis. Electrophoresis 2001;22:77 – 87.
[150] Boone CM, Waterval JC, Lingeman H, Ensing K, Underberg
WJ. Capillary electrophoresis as a versatile tool for the bio-
analysis of drugs—a review. J Pharm Biomed Anal 1999;
20:831 – 63.
[151] Boone CM, Franke JP, de Zeeuw RA, Ensing K. Intra- and
interinstrument reproducibility of migration parameters in
capillary electrophoresis for substance identification in sys-
tematic toxicological analysis. Electrophoresis 2000;21:
1545 – 51.
[152] Boone CM, Manetto G, Tagliaro F, Waterval JCM, Under-
berg WJM, Franke J-P, et al. Interlaboratory reproducibility
of mobility parameters in capillary electrophoresis for sub-
stance identification in systematic toxicological analysis.
Electrophoresis 2002;23:67 – 73.
[153] Shihabi Z. Serum drug monitoring by capillary electropho-
resis. In: Petersen JR, Mohammad AA, editors. Clinical and
forensic applications of capillary electrophoresis. Totowa,
NJ: Humana Press; 2001. p. 335 – 83.
[154] Thormann W, Caslavska J. Clinical and forensic drug toxicol-
ogy: analysis of illicit and abused drugs in urine by capillary
electrophoresis. In: Petersen JR, Mohammad AA, editors.
Clinical and forensic applications of capillary electrophoresis.
Totowa, NJ: Humana Press; 2001. p. 397 – 422.
[155] Hudson JC, Malcolm MJ, Golin M. Screening biological
specimens for drugs of forensic significance. In: Petersen
27
JR, Mohammad AA, editors. Clinical and forensic appli-
cations of capillary electrophoresis. Totowa, NJ: Humana
Press; 2001. p. 423 – 36.
[156] Wey AB, Caslavska J, Thormann W. Analysis of codeine,
dihydrocodeine and their glucuronides in human urine by
electrokinetic capillary immunoassays and capillary electro-
phoresis-ion trap mass spectrometry. J Chromatogr 2000;895:
133 – 46.
[157] Maurer HH. Systematic toxicological analysis procedures for
acidic drugs and/or metabolites relevant to clinical and for-
ensic toxicology and/or doping control. J Chromatogr B
1999;733:3 – 25.
[158] McClean S, O’Kane E, Hillis J, Smyth WF. Determination of
1,4-benzodiazepines and their metabolites by capillary elec-
trophoresis and high-performance liquid chromatography us-
ing ultraviolet and electrospray ionisation mass spectrometry.
J Chromatogr 1999;838:273 – 91.
[159] Ramseler A, Siethoff C, Caslavska J, Thormann W. Confir-
mation testing of amphetamines and designer drugs in human
urine by capillary electrophoresis-ion trap mass spectrome-
try. Electrophoresis 2000;21:380 – 7.
[160] Wey B, Thormann W. Capillary electrophoresis-electrospray
ionization ion trap mass spectrometry for analysis and con-
firmation testing of morphine and related compounds in
urine. J Chromatogr A 2002;916:225 – 38.
[161] Lurie I, Anex DS, Fintschenko Y, Choi W-Y. Profiling of
impurities in heroin by capillary electrophoresis and laser-
induced fluorescence detection. J Chromatogr A 2001;924:
421 – 7.
[162] Oda RP, Roche ME, Landers JP, Shihabi ZK. Capillary elec-
trophoresis for the analysis of drugs in biological fluids. In:
Landers JP, editor. Handbook of capillary electrophoresis.
Boca Raton, FL: CRC Press; 1997. p. 567 – 90.
[163] Taylor WJ, Diers Caviness MH. A textbook for the clinical
application of TDM. Irving, TX: Abbott Laboratories; 1986.
[164] DeCresce R, Mazura A, Lifshitz M, Tilson J. Drug testing in
the workplace. Chicago, IL: ASCP Press; 1989.
[165] Wong SHY, Sunshine I, editors. Handbook of analytical ther-
apeutic drug monitoring and toxicology. Boca Raton: CRC
Press; 1997.
[166] Binder SR. Analysis of drugs of abuse in biological fluids by
liquid chromatography. Adv Chromatogr 1996;36:201 – 71.
[167] Adams AK, Essien H, Binder SR. Identification of drugs in
physiological fluids following on-line liquid chromato-
graphic purification and analysis. Ann Biol Clin 1991;49:
291 – 7.
[168] Thormann W, Aebi Y, Lanz M, Caslavska J. Capillary electro-
phoresis in clinical toxicology. Forensic Sci Int 1998;92:
157 – 83.
[169] Thormann W, Theurillat R, Wind M, Kuldvee R. Therapeu-
tic drug monitoring of antieplieptics by capillary electropho-
resis. Characterization of assays via analysis of quality
control sera containing 14 analytes. J Chromatogr A 2001;
924:429 – 37.
[170] Cherkaoui S. Combining capillary electrophoresis with elec-
trospray ionization mass spectrometry. In: Petersen JR, Mo-
hammad AA, editors. Clinical and forensic applications of
28 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
capillary electrophoresis. Totowa, NJ: Humana Press; 2001.
p. 285 – 313.
[171] Hudson JC, Golin M, Malcolm MJ. Capillary zone electro-
phoresis in a comprehensive screen for basic drugs in whole
blood. Can Soc Forensic Sci J 1995;28:137 – 52.
[172] Hudson JC, Golin M, Malcolm MJ, Whiting CF. Capillary
zone electrophoresis in a comprehensive screen for drugs of
forensic interest in whole blood: an update. Can Soc Forensic
Sci J 1998;31:1 – 29.
[173] DeFrancesco L. Capillary electrophoresis: finding a niche.
Today’s Chem 2002;73:59 – 64.
[174] Beaubier NT, Hart AP, Bartolo C, Willman CL, Viswanatha
DS. Comparison of capillary electrophoresis and polyacryla-
mide gel electrophoresis for the evaluation of T and B cell
clonality by polymerase chain reaction. Diagn Mol Pathol
2000;9:121 – 31.
[175] Tang YW, Von Graevenitz A, Waddington MG, Hopkins
MK, Smith DH, Li H, et al. Identification of coryneform
bacterial isolates by ribosomal DNA sequences. J Clin Mi-
crobiol 2000;38:1676 – 8.
[176] Arakawa H, Watanabe K, Kashiwazaki H, Maeda M. Detec-
tion of two variant Vero toxin genes in Escherichia coli by
capillary electrophoresis. Biomed Chromatogr 2002 Feb;
16(1):41 – 6.
[177] Chen YC, Eisner JD, Kattar MM, Rassoulian-Barrett SL,
Lafe K, Bui U, et al. Polymorphic internal transcribed spacer
region 1 DNA sequences identify medically important
yeasts. J Clin Microbiol 2001;39:4042 – 51.
[178] Thomas GA, Williams DL, Soper SA. Capillary electropho-
resis-based heteroduplex analysis with a universal heterodu-
plex generator for detection of point mutations associated
with rifampin resistance in tuberculosis. Clin Chem 2001;
47:1195 – 203.
[179] Baele M, Storms V, Haesebrouck F, Devriese LA, Gillis M,
Verschraegen G, et al. Application and evaluation of the
interlaboratory reproducibility of tRNA intergenic length
polymorphism analysis (tDNA-PCR) for identification of
Streptococcus species. J Clin Microbiol 2001;39:1436 – 42.
[180] Clarridge III JE, Attorri SM, Zhang Q, Bartell J. 16S Ribo-
somal DNA Sequence analysis distinguishes biotypes of
Streptococcus bovis biotype II/2 is a separate genospecies
and the predominant isolate in adult males. J Clin Microbiol
2001;39:1549 – 52.
[181] Yu Z, Scheer WD, Hempe JM. Quantification of human
cytomegalovirus by competitive PCR and capillary electro-
phoresis. In: Palfrey SM, editor. Methods in molecular med-
icine: clinical applications of capillary electrophoresis.
Totowa, NJ: Humana Press; 1999. p. 65 – 80.
[182] Udall Jr JN, Hempe JM, Schmidt-Sommerfeld E, Scheer WD,
Mannick E, Blecker U, et al. Longitudinal analysis of plasma
cytomegalovirus deoxyribonucleic acid in a boy with Crohn’s
disease and cytomegalovirus gastroenteritis. J Pediatr Gastro-
enterol Nutr 1999;28:502 – 5.
[183] Hofgartner WT, Hühmer AFR, Landers JP, Kant JA. Rapid
diagnosis of Herpes simplex encephalitis using microchip
electrophoresis of PCR products. Clin Chem 1999;45:
2120 – 8.
[184] Merel P, Pellegrin I, Garrigue I, Caumont A, Schrive MH,
Birac V, et al. Comparison of capillary electrophoresis se-
quencing with the new CEQ 2000 DNA Analysis System to
conventional gel based systems for HIV drug resistance anal-
ysis. J Virol Methods 2001;98:9 – 16.
[185] Doglio A, Laffont C, Thyss S, Lefebvre JC. Rapid genotyp-
ing of hepatitis C virus by direct cycle sequencing of PCR-
amplified cDNAs and capillary electrophoresis analysis. Res
Virol 1998;149:219 – 27.
[186] Cornberg M, Wedemeyer H, Manns MP. Hepatitis C: ther-
apeutic perspectives. Forum (Genova) 2001;11:154 – 62.
[187] Patel JB, Leonard DG, Pan X, Musser JM, Berman RE,
Nachamkin I. Sequence based identification of Mycobacte-
rium species using the Microseq rDNA bacterial identifica-
tion system. J Clin Microbiol 2000;38:246 – 51.
[188] Jackson CJ, Barton RC, Kelly SL, Evans EG. Strain identi-
fication of Trichophyton rubrum by specific amplification of
subrepeat elements in the ribosomal DNA nontranscribed
spacer. J Clin Microbiol 2000;38:4527 – 34.
[189] Buzina W, Lang-Loidolt D, Braun H, Freudenschuss K,
Stammberger H. Development of molecular methods for
identification of Schizophyllum commune form clinical sam-
ples. J Clin Microbiol 2001;39:2391 – 6.
[190] Kurzai O, Korting HC, Harmsen D, Bautsch W, Molitor M,
Frosch M, et al. Molecular and phenotypic identification of
the yeast pathogen Candida dubliniensis. J Mol Med
2000;78:521 – 9.
[191] Payne DA, Straten MV, Carrasco D, Tyring SK. Molecular
diagnosis of skin-associated infectious agents. Arch Derma-
tol 2001;137:1497 – 502.
[192] Telatar M, Grody WW, Emmanouilides C. Detection of bcl-
2/IgH rearrangements by quantitative-competitive PCR and
capillary electrophoresis. Mol Diagn 2001;6:161 – 8.
[193] Sprouse JT, Werling R, Hanke D, Lakey C, McDonnel L,
Wood BL, et al. T-cell clonality determination using poly-
merase chain reaction (PCR) amplification of the T-cell re-
ceptor gamma-chain gene and capillary electrophoresis of
fluorescently labeled PCR products. Am J Clin Pathol 2000;
113:838 – 50.
[194] Sanchez-Vega B, Vega F, Hai S, Medeiros LJ, Luthra R. Real-
time t(14;18)(q32;q21) PCR assay combined with high-reso-
lution capillary electrophoresis: a novel and rapid approach
that allows accurate quantitation and size determination of
bcl-2/JH fusion sequences. Mod Pathol 2002;15:448 – 53.
[195] Luo V, Lessin SR, Wilson RB, Rennert H, Tozer C, Benoit B,
et al. Detection of clonal T-cell receptor gamma gene rear-
rangements using fluorescent-based PCR and automated
high-resolution capillary electrophoresis. Mol Diagn 2001;
6:169 – 79.
[196] Vega F, Medeiros LJ, Jones D, Abruzzo LV, Lai R, Manning
J, et al. A novel four-color PCR assay to assess T-cell receptor
gamma gene rearrangements in lymphoproliferative lesions.
Am J Clin Pathol 2001;116:17 – 24.
[197] Brown HA, Macon WR, Kurtin PJ, Gibson LE. Cutaneous
involvement by angioimmunoblastic T-cell lymphoma with
remarkable heterogeneous Epstein – Barr virus expression.
J Cutan Pathol 2001;28:432 – 8.
 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
[198] Welterlin V, Debecker A, Tschieb D, Zanetti C, Lange W,
Henon PR. Improvement of clonality detection rate in multi-
ple myeloma using fluorescent IgH PCR with different sets
of primers. J Hematother Stem Cell Res 2000;9:983 – 91.
[199] Lee SC, Berg KD, Racke FK, Griffin CA, Eshleman JR.
‘Pseudo-spikes’ are common in histologically benign lym-
phoid tissues. J Mol Diagn 2000;2:145 – 52.
[200] Mathew P, Hudnall SD, Elghetany T, Payne DA. T-gamma
gene rearrangement and CMV mononucleosis. Am J Hematol
2001;66:64 – 6.
[201] Berg KD, Brinster NK, Huhn KM, Goggins MG, Jones RJ,
Makary A, et al. Transmission of a T-cell lymphoma during
allogeneic bone marrow transplantation. N Engl J Med
2001;345:1458 – 63.
[202] Rolston R, Sasatomi E, Hunt J, Swalsky PA, Finkelstein SD.
Distinguishing de novo second cancer formation from tumor
recurrence: mutational fingerprinting by microdissection
genotyping. J Mol Diagn 2001:3129 – 32.
[203] Sasatomi E, Finkelstein SD, Woods JD, Bakker A, Swalsky
PA, Luketich JD, et al. Comparison of accumulated allele loss
between primary tumor and lymph node metastasis in stage II
non-small cell lung carcinoma: implications for the timing of
lymph node metastasis and prognostic value. Cancer Res
2002;1(62):2681 – 9.
[204] Sell SM, Patel S, Stracner D, Meloni A. Allelic loss analysis
by capillary electrophoresis: an accurate, automated method
for detection of deletions in solid tumors. Genet Test 2001;
5:267 – 8.
[205] Berg KD, Glaser CL, Thompson RE, Hamilton SR, Griffin
CA, Eshleman JR. Detection of microsatellite instability by
fluorescence multiplex PCR. J Mol Diagn 2000;2:20 – 8.
[206] Koul A, Willen R, Bendahl PO, Nilbert M, Borg A. Dis-
tinct sets of gene alterations in endometrial carcinoma im-
plicate alternate modes of tumorigenesis. Cancer 2002;94:
2369 – 79.
[207] Suh JH, Lim SD, Kim JC, Hong SH, Kang GH. Comparison
of clinicopathologic characteristics and genetic alterations
between microsatellite instability-positive and microsatellite
instability-negative sporadic colorectal carcinomas in pa-
tients younger than 40 years old. Dis Colon Rectum 2002;
45:219 – 28.
[208] Bandoh N, Hayashi T, Kishibe K, Takahara M, Imada M,
Nonaka S, et al. Prognostic value of p53 mutations, bax, and
spontaneous apoptosis in maxillary sinus squamous cell car-
cinoma. Cancer 2002;94:1968 – 80.
[209] Kandioler D, Zwrtek R, Ludwig C, Janschek E, Ploner M,
Hofbauer F, et al. TP53 genotype but not p53 immunohisto-
chemical result predicts response to preoperative short-term
radiotherapy in rectal cancer. Ann Surg 2002;235:493 – 8.
[210] Scappaticci FA, Marina N. New molecular targets and bio-
logical therapies in sarcomas. Cancer Treat Rev 2001;27:
317 – 26.
[211] Sheng ZM, Przygodzki RM, O’Leary TJ. Rapid screening for
KIT mutations by capillary electrophoresis. Clin Chem 2001;
47:1325 – 6.
[212] Schrag D, Kuntz KM, Garber JE, Weeks JC. Life expectancy
gains from cancer prevention strategies for women with
29
breast cancer and BRCA1 or BRCA2 mutations. JAMA J
Am Med Assoc 2000;283:617 – 24.
[213] Nicoletto MO, Donach M, De Nicolo A, Artioli G, Banna G,
Monfardini S. BRCA-1 and BRCA-2 mutations as prognos-
tic factors in clinical practice and genetic counseling. Cancer
Treat Rev 2001;27:295 – 304.
[214] Cantafora A, Blotta I, Bruzzese N, Calandra S, Bertolini S.
Rapid sizing of microsatellite alleles by gel electrophoresis
on microfabricated channels: application to the D19S394
tetranucleotide repeat for cosegregation study of familial hy-
percholesterolemia. Electrophoresis 2001;22:4012 – 5.
[215] Feolo M, Fuller TC, Taylor M, Zone JJ, Neuhausen SL. A
strategy for high throughput HLA-DQ typing. J Immunol
Methods 2001;258:65 – 71.
[216] LaFountain MJ, Schwartz MB, Svete PA, Walkinshaw MA,
Buel E. TWGDAM validation of the AmpFlSTR Profiler
Plus and AmpFlSTR COfiler STR multiplex systems using
capillary electrophoresis. J Forensic Sci 2001;46:1191 – 8.
[217] Medintz IL, Berti L, Emrich CA, Tom J, Scherer JR, Mathies
RA. Genotyping energy-transfer-cassette-labeled short-tan-
dem-repeat amplicons with capillary array electrophoresis
microchannel plates. Clin Chem 2001;47:1614 – 21.
[218] Moretti TR, Baumstark AL, Defenbaugh DA, Keys KM,
Brown AL, Budowle B. Validation of STR typing by capil-
lary electrophoresis. J Forensic Sci 2001;46:661 – 76.
[219] Larsen LA, Grønskov K, Nørgaard-Pedersen B, Brøndum-
Nielsen K, Hasholt L, Vuust J. High-throughput analysis of
Fragile X (CGG)n alleles in the normal and premutation
range by PCR amplification and automated capillary electro-
phoresis. Hum Genet 1997;100:564 – 8.
[220] Larsen LA, Christiansen M, Vuust J, Skytt Andersen P.
High-throughput single-strand conformation polymorphism
analysis by automated capillary electrophoresis: robust mul-
tiplex analysis and pattern-based identification of allelic var-
iants. Hum Mutat 1999;13:318 – 27.
[221] Butler JM, Reeder DJ. Detection of DNA polymorphisms
using PCR-RFLP and capillary electrophoresis. Methods
Mol Biol 2001;163:49 – 56.
[222] O’Connell C, Atha D, Jakupciak J, Amos J, Richie K. Stand-
ardization of PCR amplification for fragile X trinucleotide
repeat measurements. Clin Genet 2002;61:13 – 20.
[223] Gomez-Llorente MA, Suarez A, Gomez-Llorente C, Munoz
A, Arauzo M, Antunez A, et al. Analysis of 31 CFTR muta-
tions in 55 families from the South of Spain. Early Hum Dev
2001;65:S161 – 4.
[224] Guttman A, Gao HG, Haas R. Rapid analysis of mitochon-
drial DNA heteroplasmy in diabetes by gel-microchip elec-
trophoresis. Clin Chem 2001;47:1469 – 72.
[225] Tang T, Badal MY, Ocvirk G, Lee WE, Bader DE, Bekkaoui
F, et al. Integrated microfluidic electrophoresis system for
analysis of genetic materials using signal amplification
methods. Anal Chem 2002;74:725 – 33.
[226] Sung WC, Lee GB, Tzeng CC, Chen SH. Plastic microchip
electrophoresis for genetic screening: the analysis of poly-
merase chain reactions products of fragile X (CGG)n alleles.
Electrophoresis 2001;22:1188 – 93.
[227] Larsen LA, Johnson M, Brown C, Christiansen M, Frank-
30 J.R. Petersen et al. / Clinica Chimica Acta 330 (2003) 1–30
Hansen R, Vuust J, et al. Automated mutation screening using
dideoxy fingerprinting and capillary array. Hum Mutat 2001;
18:451 – 7.
[228] Dorschner MO, Barden D, Stephens K. Diagnosis of five
spinocerebellar ataxia disorders by multiplex amplification
and capillary electrophoresis. J Mol Diagn 2002;4:108 – 13.
[229] Grody WW, Cutting GR, Klinger KW, Richards CS, Watson
MS, Desnick RJ. Laboratory standards and guidelines for
population-based cystic fibrosis carrier screening. Genet
Med 2001;3:149 – 54.
[230] Zabzdyr JL, Lillard SJ. Measurement of single-cell gene ex-
pression using capillary electrophoresis. Anal Chem 2001;
73:5771 – 5.
[231] Rajevic U, Juvan R, Gazvoda B, Repse S, Komel R. Assess-
ment of differential expression of oncogenes in gastric adeno-
carcinoma by fluorescent multiplex RT-PCR assay. Pflugers
Arch 2001;442:R190 – 2.
[232] Crawford EL, Peters GJ, Noordhuis P, Rots MG, Vondracek
M, Grafstrom RC, et al. Reproducible gene expression meas-
urement among multiple laboratories obtained in a blinded
study using standardized RT (StaRT)-PCR. Mol Diagn
2001;6:217 – 25.
[233] Crawford EL, Khuder SA, Durham SJ, Frampton M, Utell M,
Thilly WG, et al. Normal bronchial epithelial cell expression
of glutathione transferase P1, glutathione transferase M3, and
glutathione peroxidase is low in subjects with bronchogenic
carcinoma. Cancer Res 2000;15(60):1609 – 18.
[234] Willey JC, Crawford EL, Jackson CM, Weaver DA, Hoban
JC, Khuder SA, et al. Expression measurement of many
genes simultaneously by quantitative RT-PCR using stand-
ardized mixtures of competitive templates. Am J Respir Cell
Mol Biol 1998;19:6 – 17.
[235] Stanta G, Bonin S, Lugli M. Quantitative RT-PCR from fixed
paraffin-embedded tissues by capillary electrophoresis. Meth-
ods Mol Biol 2001;163:253 – 8.
[236] Liu Y, Ganser D, Schneider A, Liu R, Grodzinski P, Kroutch-
inina N. Microfabricated polycarbonate CE devices for DNA
analysis. Anal Chem 2001;73:4196 – 201.
[237] Nagai H, Murakami Y, Yokoyama K, Tamiya E. High-
throughput PCR in silicon based microchamber array. Bio-
sens Bioelectron 2001;16:1015 – 9.