FUNCTIONAL BIOCHEMISTRY
LESSON № 1 BIOCHEMISTRY OF BLOOD. BLOOD PLASMA PROTEINS.
Study questions.
1. Blood: composition, volume, physical and chemical characteristics of
blood (pH, osmotic pressure, specific weight). What is plasma and what
is serum?
2. List functions of blood.
3. The total blood protein: normal value, the principal of determination,
hyper- and hypoproteinemia, clinical importance of determination of the
total blood protein.
4. Separation of blood plasma proteins: salting out, electrophoresis. The
technic of the agar gel (paper) electrophoresis (short description). Blood
proteinogram and clinical importance of proteinogram. What
abnormalities can be identified?
5. Plasma albumins. Specific properties, biosynthesis, functions of plasma
albumins.
6. Functional classification of plasma proteins: transport proteins,
antiproteases (inhibitors of proteolytic enzymes) and acute phase proteins
(APP). Fill up the following table: give the brief description of the most
important representative of α1-, α2-, β- and γ-globulins
α1-globulins
α2- globulins
β- globulins
γ - globulins
7. Characteristics of plasma transport proteins (transferrin, retinol binding
protein, ceruloplasmin, transcortin, haptoglobin, hemopexin). The use in
clinical diagnostics.
8. Plasma lipoproteins. Structure (draw the coomon structure of the
lipoprotein particle), composition, specific apoproteins. Function of
Chylomicrons, VLDL, LDL and HDL.
Fill up the following table
Class of plasma Protein (%) TAG
lipoprotein and its and
the (%)
function
main
apoproteins
Chylomicron
Cholesterol Phospholipids Size
and
cholesterol
esters (%)
VLDL
LDL
HDL
9. Antiproteases: α1-antitrypsin, α2-macroglobulin, α1-antichymotrypsin.
10. Acute phase proteins as a diagnostic tool. Positive and negative acute
phase proteins.
11. Blood enzymes : secretory, indicatory and excretory enzymes.
12. Immunoglobulin’s. Structure (draw the common structure of the Ig G) ,
classes, functions, content on blood).
13. Non-protein components of plasma. Diagnostic importance of their
determination.
LABORATORY WORK
ALBUMIN ASSAY
Principle: The assay is based on the selective interaction between albumin
and Bromocresol-Green reagent causing the formation of a colored product that is
indicated at the wave length 625 nm.
Procedure: Place into the tube 0,1 ml of serum (diluted 1:10 with 0,9 %
solution of NaCl), 4,0 ml of working reagent, mix the content of the tube. After 10
min incubation read optic density at 625 nm (red light) against working reagent.
Perform the same procedure for the calibrator (albumin concentration 50 g per
liter).
Calculation:
C=Dser/Dcalib×50=
Normal values: 40-50 g/L
Result:
Resume: