CLINICAL

CHEMISTRY
P R E PA R E D B Y : C H A R I Z Z E M O N I Q U E V. D A R A U G , R M T
 PROTEINS
• Proteins are macromolecules made of amino • Structures:
acids and are mostly synthesized in the liver 1. Primary – linear sequence of the amino
and are secreted by the hepatocytes into the acid.
circulation.
2. Secondary – specific 3-D conformation
• Functions:
3. Tertiary – actual 3-D configuration ;
1.Repair body tissues affected by denaturation
2.Important in blood coagulation and 4. Quaternary – association of 2 or more
immunologic function polypeptide chains to form functional
3.For transport of metabolic substances protein molecule.
4.Maintenance of osmotic pressure – Classification
5.Buffer 1. Simple proteins
6.Biocatalysts 2. Conjugated proteins
(apoprotein + prosthetic group)
 MAJOR PLASMA PROTEINS
PREALBUMIN
ALBUMIN
ALPHA₁ GLOBULIN • α₁-antitrypsin
• AFP
• α₁-lipoprotein
• α₁-acid glycoprotein
• α₁-antichymotrypsin
• inter-alphatrypsin inhibitor
• Gc-globulin

ALPHA₂ GLOBULIN “CHA” • Ceruloplasmin

• Haptoglobin
• α₂-macroglobulin
BETA GLOBULIN “Better Complement Her Creepy • Complement
Transformation” • Hemopexin
• CRP
• Transferrin

• *Fibrinogen,VLDL, LDL, beta2 microglobulin

GAMMA GLOBULIN Immunoglobulins (IgG, IgM, IgA, IgE, IgD)

 OTHER NAME/
PROTEIN ABBREVIATION FUNCTION NOTES

Used to confirm if the specimen is

• Carrier protein for really CSF.
PREALBUMIN Transthyretin thyroid hormones  Increased: alcoholism, chronic renal failure,
and Vitamin A. steroid treatment.
 Decreased: liver disorders, inflammation,
malignancy and poor nutrition.

 Found in highest concentration in the plasma

• Serves as transport  Lifespan: 20 days
protein (Bilirubin,  Indicator of nutritional status.
ALBUMIN -
steroids, ions, fatty Most anodal protein.
acids and drugs)  Negative APR –decreases during inflammation.
• Maintains oncotic  Increased: Dehydration, prolonged
pressure.
tourniquet application or specimen
evaporation
 OTHER NAME/
PROTEIN ABBREVIATION FUNCTION NOTES

 MAJOR inhibitor of protease activity.

α₁-Antitrypsin AAT
• Neutralizes trypsin  Most abundant of the alpha1-globulin band
like enzyme  Deficiency: Emphysematous pulmonary disease
and juvenile hepatic cirrhosis

 Most abundant protein in fetal serum

 Oncofetal tumor marker found in patients with
hepatocellular carcinoma and germ cell tumor
 Specimen: Maternal serum (screening)
α₁-fetoprotein AFP Amniotic fluid (confirmatory)
 Increased: Neural tube defects, Twins, spina
bifida and fetal distress.
 Decreased: Down Syndrome (Trisomy 21),
Trisomy 18
 OTHER NAME/
PROTEIN ABBREVIATION FUNCTION NOTES

 Has greatest affinity for progesterone.

• Carrier for steroid  Useful diagnostic tool for neonates with
α₁-Acid Orosomucoid hormones bacterial meningitis.
glycoprotein (AAG) • Binds to basic drugs  Increased: RA, Pneumonia, cell
(quinidine) proliferation, pregnancy
 Decreased: nephrotic syndrome

• Binds and
inactivates PSA  Associated with the pathogenesis of
• Inhibits the activity Alzheimer’s disease.
α₁-
α₁-x of cathepsin G,  Increased: infection, malignancy, burn, AMI,
Antichymotrypsin
pancreatic elastase, and Alzheimer’s disease.
mast cell chymase  Deficiency: Liver disease
and chymotrypsin.
 OTHER
NAME/
PROTEIN ABBRE FUNCTION NOTES
VIATION

 Used to evaluate rheumatic diseases.

 Evaluates the degree of intravascular hemolysis.
• Binds FREE  Increased: inflammatory conditions, burns,
Haptoglobin - HEMOGLOBI trauma
N  Decreased: Intravascular hemolysis,
hemoglobinuria
 Methods: RID & Immunonephelometry

Haptoglobin Retics count RBC count Hgb Hct

1. Hemolytic anemia ↓ ↑ ↓ ↓ ↓
2. RBC lysis (spleen and
N ↑ - - -
liver)
3. Anemia (not related to
N ↓ - - -
RBC lysis)
4. Hepatic dysfunction
↓ - - - -
(synthesis)
 OTHER NAME/
PROTEIN ABBREVIATION FUNCTION NOTES

 Marker for Wilson’s disease

 deposition of copper in the skin, liver, brain
and cornea [Kayser-Fleisher rings]
 ↓ serum copper ↑ urinary copper

Ceruloplasmin - • Binds copper  Method: Copper oxidase activity

 Increased: inflammation, cancer,
pregnancy
 Decreased: Wilson’s disease, malnutrition,
malabsorption, nephritic disease and
Menkes’ kinky hair syndrome
 OTHER NAME/
PROTEIN ABBREVIATION
FUNCTION NOTES

 Largest major non-immunoglobulin

proteins in plasma.
 Increased: nephrotic syndrome
• Inhibits proteases
(trypsin, pepsin,
contraceptive use, pregnancy, estrogen
α₂-macroglobulin AMG therapy, diabetes and liver disease.
kallikrein, thrombin
and plasmin)  Decreased: acute inflammatory disorders
(slightly) and acute pancreatitis (markedly)
 Methods: RID,ELISA, Latex agglutination,
and Immunonephelometry.

 Promising renal marker

• Needed for CD8  Light chain component of the HLA
β₂-macroglobulin
production.  Increased: SLE, HIV, Renal failure, Multiple
myeloma, and RA.
 OTHER NAME/
PROTEIN ABBREVIATION FUNCTION NOTES

• Transports iron
 Used to determine the cause of anemia, to
• Prevents the loss of
Transferrin Siderophilin measure iron metabolism and determine
iron through the
kidney. the iron-carrying capacity of the blood.

Transferrin
Serum Iron TIBC
saturation

Iron deficiency ↓ ↓ ↑

Iron overload ↑ ↑ ↓/N

 OTHER NAME/
PROTEIN ABBREVIATION
FUNCTION NOTES

 C3- most abundant form in serum

• Protects human  Increased: inflammatory conditions
Complement -
body from infection  Decreased: DIC, hemolytic anemia and
malnutrition
 Serves as non-specific indicator of
inflammation.
 General scavenger molecule
 In cases of SLE & RA – used to assess
C-reactive protein CRP • Cardiac marker
effectiveness of a specific arthritis
treatment.
 Increased: tissue necrosis, rheumatic
fever, infections, MI, RA and gout.

 Has the strongest affinity for heme.

Hemopexin • Binds HEME  Increased: inflammation, DM, melanomas,
Duchenne’s type muscular dystrophy.
 OTHER NAME/
PROTEIN ABBREVIATION
FUNCTION NOTES

 Synthesized by the plasma cells.

 IgG – crosses placenta; most abundant
 IgA- found in secretions
 IgM – first to increase in response to antigenic
• Humoral
Immunoglobulins Antibodies stimulations
immunity
 IgD – present mostly on the surface of B cells
 IgE – associated with allergic and anaphylactic
response.

 Present in plasma but not in serum.

 Most abundant of the coagulation factor.
 May serve as marker for long-term prognosis of
• For
Fibrinogen cardiovascular disease.
coagulation
 Increased: inflammatory disorders, pregnancy
and use of oral contraceptives.
 Decreased: extensive coagulation.
 ACUTE PHASE REACTANTS
• AAT
• α₁-acid- glycoprotein
• Ceruloplasmin
• Complement proteins
• CRP
• Factor VIII
• Fibrinogen A2C3F3HaILSAT
• Ferritin
• Haptoglobin
• Immunoglobulins
• Lipoproteins
• Serum amyloid protein A
• Albumin (Negative APR)
• Transferrin (Negative APR)
 MISCELLANEOUS PROTEINS
1.Bence-Jones proteins
– Identical light chains (kappa-kappa or lambda-lambda)
– Precipitates at 40-60˚C but dissolves at 100˚C
– Excreted by the patients with multiple myeloma.
– Method: Immunofixation

2.Myoglobin (Screening) Rises: 1-3 hours

Peaks: 5-12 hours
– Earliest cardiac marker but not specific for AMI. Normalize: 18-30 hours
– Small heme protein found in skeletal and cardiac marker (Rodriguez)
– Has higher affinity for oxygen than hemoglobin
Rises: 2-3 hours
– Increased: AMI, Angina, rhabdomyolysis, muscle trauma, Peaks: 8-12 hours
extraneous exercise, intramuscular injection and acute renal Normalize: 18-30 hours
failure. (Bishop)
MISCELLANEOUS PROTEINS
3.Troponin (Confirmatory)
– Complex of 3 proteins:
a. TnI (inhibitory subunit) Rises: 3-6 hours
Peaks: 12-18 hours or 14-20 hours
- Found in the MYOCARDIUM Normalize: 5-10days

- HIGHLY SPECIFIC for AMI

b. TnT (tropomyosin-binding subunit) Rises: 3-4 hours
- SENSITIVE MARKER for diagnosis of angina at rest Peaks: 10-24 hours
Normalize: 7 days
- Useful for assessment of early and late AMI (but may remain for
10-14 days)
c. TnC (calcium-binding subunit)
– TnI and TnT are specific for heart muscles and are almost
absent in normal serum.
MISCELLANEOUS PROTEINS
4.B-type natriuretic peptide (BNP)
– Diagnostic of congestive heart failure.
5.Cystatin C
– Low molecular weight protein
– Cysteine proteinase inhibitor
– Recently proposed new marker for early assessment of changes to the
GFR.
– Increased: Renal disease
– Method: particle-enhanced immunoturbidimetry, immunonephelometry
URINARY PROTEINS
• Normal bilirubin excretion rate: <20ug/min
• Urine protein of ≥6 mg/dL produces color change on urine dipstick.
• Proteinuria (>0.5mg/dL)
1. Glomerular proteinuria (Albuminuria) – most common and serious type.
2. Tubular proteinuria – due to defective reabsorption
3. Overload proteinuria – includes hemoglobinuria, myoglobinuria and Bence-Jones
proteins
4. Post-renal proteinuria – proteins coming from the urinary tract caused by infection,
bleeding or malignancy.
• Microalbuminuria
- Early indicator of glomerular dysfunction and precedes to diabetic nephropathy
- Specimen: Random urine Reference value: 0-29ug/mg crea
- Method: random-spot albumin-creatinine ratio Microalbuminuria: 30-300ug/mg crea
Clinical albuminuria: >300ug/mg crea
CSF PROTEINS
• Protein in CSF is less than 1% compared to plasma protein
• Method: TCA, SSA, Coomassie Brilliant Blue, Lowry and Kinetic Biuret reaction
• Reference value: 15-45mg/dL
• Increased: meningitis, traumatic tap, multiple sclerosis, intracerebral hemorrhage,
myxedema, drug toxicity
• Decreased: intracranial hypertension, hyperthyroidism, leakage of CSF due to trauma

CSF OLIGOCLONAL BANDS

(MS NENG) SERUM OLIGOCLONAL BANDS
• Multiple Sclerosis (VLL)
• Neurosyphilis • Viral infections
• Encephalitis • Leukemia
• Neoplastic disorders • lymphoma
• Guillain-Barre Syndrome
AMINOACIDOPATHIES
1. ALKAPTONURIA
• Absence of homogentisate-oxidase in the tyrosine pathway.
• Darkening of urine upon standing at room temperature.
• Clinical feature: Onchronosis (tissue pigmentation)
2.HOMOCYSTINURIA
• Impaired activity of cystathionine B-synthetase
• Elevated levels of homocysteine and methionine in plasma and urine.
• Screening test: Modified Guthrie Test (Antagonist: L-methionine sulfoxime)
3.MAPLE SYRUP URINE DISEASE (MSUD)
• Markedly reduced or absence of a-ketoacid decarboxylase.
• Accumulation of Leucine, Isoleucine and Valine in blood, urine and CSF.
• Causes mental retardation or death in infants.
• Screening test: Modified Guthrie Test (Antagonist: 4-azaleucine)
• Diagnostic: Amino acid analysis (HPLC)
• Indicator: 4mg/dL of leucine is indicative of MSUD
 AMINOACIDOPATHIES
4.PHENYLKETONURIA (PKU)
• Deficiency of phenylalanine hydrolase and also tetrahydrobiopterin.
• “Musty” odor of urine
• Causes mental retardation in children.
• Screening test: Modified Guthrie Inhibition Assay (B.subtilis and β₂-thienylalanine) (+)
result: bacterial growth if the phenylalanine is >4mg/dL
5.TYROSINEMIA
• Deficiency of either tyrosine aminotransferase; 4-hydroxyphenypyruvic acid oxidase;
fumarylacetoacetate FAA hydrolase