Electophoresis

Question 1

Principle of electrophoresis

Answer 1

most frequency used in clincal practice, represents a method of seperartion of charged particles based on their different movement when they are placed in an electrical filed.

Charged particles move toword the opossite charged electrode:

• cations (positively charged particles) move to the catode (negatively charged electrode
• anions (negatively charged particles) move to the anode (positively charged electrode)

Question 2

The rate of migration in electrophoresis depends on:

Answer 2

- the size of the particles (bigger particles move slower)

- the shape of the particels

• the net charge of the particles (more negatively charged ions move quicker)
• the mass of the particles
• electrical force applied on the charged particles
• the characteristics of the supporting medium and the properties of the buffer

supporting material: filter paper or sheets (Blatt) of plastik like cellulose acetate membranes, polyacrylamide, starch, agar or agarose gels)

properties of the buffer: pH, ionic strength, compositon

Question 3

Immunglobulin G (IgG)

Answer 3

• is the prototype of the immunglobuline family
• the most abudant antibody in plasma (represents about 75% of total immunglobuline in serum)
• major antibody of the secondary immunse response (the synthesis takes a longer period than that of IgM)
• can cross the placenta (major protective antibody in newborns)

Question 4

Immunglobulin M (IgM)

Answer 4

• is the first immunglobulin produced in response to an antigen

Question 5

Immunglobulin A (IgA)

Answer 5

• is the major antibody in external secretions like salvia, tears, sweat, bronchial mucus intestinal mucus

Question 6

Immunglobulin D (IgD)

Answer 6

• has an unknwon function

Question 7

Immunglobulin E (IgE)

Answer 7

• plays am important role in allergic reactions

Question 8

Total serum protein in g/dl + seperated in albumin and globulin

Answer 8

Total serum protein: 6 - 8 g/dl;

albumin: 4,5 - 5 g/dl

globulin: 1,5 - 2,5 g/dl

Question 9

Pathological modification is case of

acute inflammation

Answer 9

↑ α1-globulins

↑ α2-globulins

Question 10

Pathological modifications in case of

chronic inflammation

Answer 10

↑ γ-globulins

Question 11

Pathological modifications in case of

multiple myeloma

Answer 11

↑ Total proteins

↑ γ-globulins

Question 12

Pathological conditions in case of

hepatic cirrhosis

Answer 12

↓ Total proteins

↓ Albumin

↑ β-globulins

↑ γ-globulins

Question 13

Patological modifications in the case of

nephrotic syndrome

Answer 13

↓ Total proteins

↓ Albumin

↑ α2-globulins
↑ β-globulins
↓ γ-globulins (in severe forms)

Question 14

Pathological modifications in the case of

hypogammaglobulinemia

Answer 14

↓Totalproteins

↓ γ-globulins

Question 15

Choose the correct answer: In acute inflammation are increased:

a. α1- and γ-globulins;
b. α2- and γ-globulins;
c. β- and γ-globulins;
d. γ-globulins;
e. α1- and α2-globulins

Answer 15

e. α1- and α2-globulins

Question 16

Complete the blank spaces with one of the following words: increased, decreased, unmodified.

In nephrotic syndrome:

albumin is…..,

α1-globulins are…..,

α2-globulins are…..,

β- globulins are…..

γ-globulins are…..

Answer 16

decreased

unmodified

increased

increased

decreased

Question 17

Specify the name of the disease you suppose to be corresponding to the following results of protein electrophoresis. Explain your answer.

Albumin: 45%
α1-Globulins: 5%

α2-Globulins: 8%

β-Globulins: 17%

γ-Globulins: 25%

Total serum proteins: 5.4g/dl

Answer 17

Hepatic cirrhosis

Question 18

Normal aspect of human serum electrophoresis

Answer 18

Question 19

Albumin

Answer 19

• the major protein fraction in serum (represent 52-59% of total serum proteins)
• synthesized by the liver as a single polypeptide chain (with 585 amino acids and MW of 65000 daltons)
• role: maintaning the osmotic pressure of the blood and to transport small molecules like ions (Ca, Cu, heavy metals), bilirubin, fatty acids, hormons, certain drugs (salicylates like aspirin and dicoumarol)

Question 20

Globulin

Answer 20

• divided into four fractions:

albumin, alpha1 (α1), alpha2 (α2), beta (β) and gamma (γ) globulins

each fraction contains different proteins with specific function

Question 21

α1-Globulins

Answer 21

Question 22

α2-Globulins

Answer 22

Question 23

β-Globulins

Answer 23

Question 24

γ-Globulins

Answer 24

● Immunoglobulin G (IgG)

● Immunoglobulin M (IgM)

● Immunoglobulin A (IgA)

● Immunoglobulin E (IgE)

● Immunoglobulin D (IgD)

Question 25

Hypoproteinemia

Answer 25

liver diseases (synthesis of proteins is affected): heptaic cirrhosis, chronic hepatitis, liver tumors

kidney diseases (proteins are lost through the kidneys): glomerulonephritis, nephrotic syndrome

Question 26

Hyperproteinemia

Answer 26

monoclonal gammapathies: multiple myelom, Wadenstrom macroglobulinemia etc.

Question 27

Protein electrophoresis

Answer 27

a protein is a charged particle (ampholyte) whose net charge can be either positive or negative

If the buffer has pH < pH_i of the ampholyte => it binds H+ ions become positively charged (move to katode)

If the buffer has pH > pH_i of the amphoyte => it losses H+ ions become negatively charged (move to anode)

A particle without net charge is not migrating in an electrical field and remains at the point of application,

Question 28

Seperation of serum proteins take plase..

Answer 28

in an alkaline pH (8,5-8,6), higher than the pHi

at this pH the net charge of the proteins are negative and they will move towards the anode

y globulin migrate the least, albumin is the farhtest !