Clinical Chemistry

Recommended routine method for determining total protein

Colorimetry –> Biuret technique

Alkaline medium: Copper salts + protein –> purple complex @ 540nm

Protein – Spectrophotometry

UV light absorbance @?

210 & 280nm
Serum proteins with net negative charge

gamma globulins

endosmotic force–>cathode (-)

High Resolution Serum Protein Electrophoresis

Proteins?

Albumin – a1 – a2 – beta – gamma

Two peaks are seen in albumin band

Dx?

Bisalbuminemia<– variant Albumin allotype

AFP

migrates in ? band on electrophoresis

a1
Clinical utility of albumin

nutritional status–> t1/2= 17d

hepatic synthetic function–> end stage liver disease

diabetic control–>nonenzymatic glycosylation (nml<8%)

Albumin = negative acute phase reactant

 

Prealbumin

Electrophoresis and function

faint–> only seeen on HR SPEP

aka transthyretin–> binds T3/4, VitA complex

Amyloid precursor protein in senile cardiac amyloidosis
Prealbumin
Protein responsible for familial amyloid polyneuropathy
mutant versions of tranthyretin/prealbumin
Clinical utility of prealbumin

Nutritional status –> t1/2 = 48hrs

CSF PEP–> sharp prealbumin band

SPEP: prominent prealbumin band

DDx?

Pt on heparin=> alteration in b-lipoprotein

Chronic alcoholic

Corticosteroid Rx

Major component of a1-band
a1-antitrypsin –> + acute phase reactant

Acute inflammation

Major component of increased a1-band

a1-acid glycoprotein (orosomucoid)

~ monitoring UC

Elevated a2-macroglobulin

DDx?

Liver / renal disease

decreased Ceruloplasmin

DDx?

= a2-band, Wilson’s disease

hepatic failure, malnutrition, Menke syndrome

decreased haptoglobin

Dx?

a2-band, acute phase reactant

intravascular hemolysis

Independent risk factor for CVD in diabetics?
Haptoglobin, phenotype 2-2
b-globulins

Transferrin–>major protein

Fibrinogen–> normal = no fibrinogen in serum

~seen in heparinized patient<–incomplete blood clot

Complement (C3)

Increased Transferrin

DDx?

Iron deficiency–>marked –>ddx abnormal band–> M-protein

Pregnancy

Estrogen Rx

SPEP: band in b-g interface

which protein? how  do you obviate this interference?

Fibrinogen

Absolute Ethanol

Protein that can be misinterpreted as an IgM in serum

Causes?

Fibrinogen

dyfibrinogenemia, APL syndrome, liver dz,

VitK deficiency,heparin

g- region proteins

Ig and CRP

“C”= “gamma”

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