C H A P T E R
Phadia Laboratory Systems
7.19 Angela Gore1 (angela.gore@thermofisher.com)
Gareth Evans2
Mats Rilvën2
The Phadia Laboratory Systems (formerly known as Uni-
CAP®, then latterly ImmunoCAP®) include the Phadia
100, Phadia 250, Phadia 1000, Phadia 2500, and Phadia
5000. They are all fully integrated laboratory systems
designed for the testing of allergy and autoimmune dis-
eases. The reagents for the allergy assays are based on the
well-established ImmunoCAP format, whereas the EliA®
autoimmunity assays use normal ELISA-type-coated wells,
although both assay types can be run together in the same
run (with the exception of the Phadia 100 and Phadia
1000). All the instruments provide full automation and
user-friendly software and deliver fast and accurate test
results that provide physicians with valuable clinical infor-
mation. Over 500 different allergens and allergen compo-
nents are available for allergy testing, and 32 different
autoimmune markers, with new analytes continually being
developed and released for both disease areas. The Phadia
Laboratory Systems and reagents have been developed by
Phadia, the world leader in allergy and autoimmunity
diagnostics, which is now the ImmunoDiagnostics busi-
ness of Thermo Fisher Scientific.
The Phadia Laboratory Systems’ group of instruments
ranges from the benchtop Phadia 100, which is a batch pro-
cessor capable of producing up to 46 results in one run tak-
ing about 2.5 h, through to the Phadia 5000 which can
produce up to 5000 results per working day. All instruments
are fully automated, incorporating sample and reagent han-
dling, washing, reading, calculation, and integrated pro-
cessing and are all controlled by the ImmunoCAP Data
Manager (IDM) software which links to laboratory infor-
mation management systems (LIMSs) (Fig. 1).
Typical Assay Protocol
Assay protocols are similar on all instruments, however, on
the larger instruments, processing time is less due to dif-
ferences in handling of individual wells and samples. In the
first step, sample, ImmunoCAP or EliA well, and reagent
are dispensed. There is an incubation, followed by wash-
ing, rinsing, and aspiration. The signal generation stage
consists of substrate addition, stop reagent addition, and
fluorescent signal measurement. On the Phadia 100,
reagents are loaded for each run, whereas on the larger
instruments, all reagents are stored onboard, and continu-
ous loading of samples is possible.
1This edition.
2Previous editions.
© 2013 David G. Wild. Published by Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/B978-0-08-097037-0.00052-X
The systems are designed to use stored calibration
curves, with a new curve required approximately once a
month or with every new lot number of conjugate. Between
calibrations, curve controls are used to check performance
against the stored curve.
Results can be exported to LIMS as soon as they are
available on the larger instruments, and laboratory reports
can be printed via IDM when the process is ended.
Assay Principle
See Fig. 2.
IMMUNOCAP ALLERGY ASSAYS
The Phadia Laboratory Systems’ allergy assays are built
around a solid phase consisting of a flexible hydrophilic
carrier polymer encased in a capsule—ImmunoCAP. The
carrier consists of a cyanogen bromide (CNBr)-activated
cellulose derivative that can bind at least three times more
protein than a paper disc and about 150 times more than
can be adsorbed to the inner surface of a traditional coated
tube or microtiter well (Fig. 3). This solid phase is an
excellent carrier of antigens (allergens) and provides
favorable reaction conditions, including short diffusion
distances.
The allergens coated onto the ImmunoCAP can be
either recombinant or native whole allergen extracts,
either for a single allergen or a mixture of allergens. Also,
now molecular allergology is available, where single indi-
vidual allergen components are used to quantitatively
detect IgE antibodies. This gives added information to cli-
nicians in terms of assessing the risk of allergic reaction,
identifying cross-reactivity, and determining a patient’s
suitability for specific immunotherapy.
ELIA AUTOIMMUNITY ASSAYS
The ImmunoCAP system is ideal for allergy assays as it
allows an excess of allergen to be bound to the solid phase.
However, this is not required for autoimmunity assays, so
the EliA system uses conventional individual microtiter
wells for antigen coating. The antigens bound to this solid
phase are mainly human recombinant, produced in the
Baculovirus/Insect Cell System. This method of antigen
production ensures antigens of constant high purity, sta-
bility, and quality, which have the correct sequence and
structure, and can also be produced in very large quanti-
ties, which in turn minimises any lot-to-lot variances.
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