Aspek Laboratorium

Pemeriksaan Protein
Ariosta Setyadi
11/25/2017 2
 Serum Protein Electrophoresis
(SPE)
Electrophoresis is refer to
the migration of all charged
solutes or particle in liquid
medium under influence of
an electric field

3
 http://erl.pathology.iupui.edu/
Generator.cfm?Image=SPE_16

Normal pattern
Albumin 47-71% (3.63-4.91 g/dL)
Alpha-1 globumin 2.7-5.8% (0.11-0.35 g/dL)
Alpha-2 globulin 5.1-12.0% (0.65-1.17 g/dL)
Beta- globulin 4.5-15.7% (0.74-1.26 g/dL)
Gamma globulin 11.3-24.0% (0.58-1.74 g/dL)

4
 Plasma proteins
Total protein
test performed on serum (no fibrinogen)
Colorimetric method based on purple color
development in alkaline solutions of copper
sulfate in the presence of substances with two
or more peptide bonds
normal range 6.5 - 8.5g/dl
sensitivity to 10 - 15 mg/dl

5
 Total protein measurement
Method How it works Sensitivity Range
Biuret Cu2+-peptide 1-10 mg protein
bond complex
Lowry Heavy metal complex 20-300 mg protein
with aromatic amino acid
Bradford Dye reaction with 10-100 mg protein
amino group side
chain (Lys)

6
Clinical significance of Total
protein
Use - screen for nutritional deficiencies and
gammopathies
Increased in hypergammaglobulinemia poly or
monoclonal
decreased
in nutritional deficiencies
decreased production or increased loss or increased
catabolism
dilutional (iv fluids)
7
 Albumin measurement
Albumin is measured
spectrophotometrically following specific
binding to certain dyes (bromcresol green
or bromocresol purple)

Albumin + BCG Albumin-BCG complex

8
 Clinical Significance of
Albumin
Reference range (3.5-5.5 g/dL)
Hyperalbuminemia dehydration
Hypoalbuminemia
Low intake, synthesis
Increase loss:
kidney; nephrotic syndrome, wound, burn
GI tract; protein-losing enteropathy

9
 Total Globulin
Total globulin is a calculated value of
the difference between total protein
and albumin

Globulin = Total protein - Albumin

normal values 2-3.5 g/dl

10
 A/G RATIO
[albumin]
[TP] - [albumin]
Reference Range: 1.6-2.0:1
Abnormal Values:
Increased Values: hypogammaglobulinemia; etc.
Decreased Values: infections; renal disease; liver disorders
The value in determining the A/G ratio lies in those cases where
changes in the protein values are associated with water imbalances
rather than protein problems.
Generally this is associated with dehydration, which would yield an
increased TP and albumin, but the A/G ratio would be normal,
indicating that the protein levels are actually normal
11
 Reference Values

Total protein 6.0-8.0 g/dL

12
 Serum Protein Electrophoresis (SPE)

http://erl.pathology.iupui.edu/
Generator.cfm?Image=SPE_16

Normal pattern
Albumin 47-71% (3.63-4.91 g/dL)
Alpha-1 globumin 2.7-5.8% (0.11-0.35 g/dL)
Alpha-2 globulin 5.1-12.0% (0.65-1.17 g/dL)
Beta- globulin 4.5-15.7% (0.74-1.26 g/dL)
Gamma globulin 11.3-24.0% (0.58-1.74 g/dL)
13
 Normal serum protein
electrophoresis
Albumin migrates most to the anode
The five regions are defined
Albumin
alpha-1 globulin
alpha-2 globulin
beta-globulin
gamma-globulin
Gamma-globulins are dragged to the cathode
despite slight negative charge

14
 Specific Proteins in Plasma (1)

Pre albumin region

15
 Specific Proteins in Plasma (2)

Albumin region
Albumin
Most abundant protein in the serum 3.5 - 4.5g/dl or
35 - 45g/L
possess low pI, thus confer anodal migration
Critical role in maintaining oncotic pressure
(decrease in Alb = edema)
Carrier protein for many ligands
Source of amino acids from the liver to cells

16
 Specific Proteins in Plasma (3)
a1-region
Alpha-1 antitrypsin (AAT) is the major protein in this
region
modulates endogenous proteolysis (trypsin and other
proteases)
Normal allele M . Homozygous for an abnormal ZZ
allele develop early pulmonary emphysema and
cirrhosis
AAT is an acute phase reactant increasing in acute
inflammation

17
 Normal

Alpha-1-anti-trypsin deficiency pattern

http://erl.pathology.iupui.edu/labmed/Generator.cfm?Image=SPE
18
Specific Proteins in Plasma (4)

a2 globulins
a2 macroglobulin (AMG), also a protease
inhibitor is very large
In nephrotic syndrome , smaller proteins are lost in
urine and AMG increases 10X maintaining the
oncotic pressure The AMC reaches levels similar to
albumin

19
 Normal

The nephrotic pattern

http://erl.pathology.iupui.edu/labmed/Generator.cfm?Image=SPE
20
Specific Proteins in Plasma (5)

a2 globulins (2)
Haptoglobin combines with Hb to preserve
iron. Removed immediately by RES
Ceruloplasmin
Facilitate oxidation-reduction, which is
important in mediating binding between
iron and transferrin
Decrease in Wilson disease (autosomal
recessive that reduced intracellular ATPase
activity)

21
Specific Proteins in Plasma (6)

Beta globulins
Transferin transports ferric iron from
intracellular ferritin to bone marrow
erythroid precursors have receptors
Fibrinogen coagulation factor
fibrin monomers polymerize to form
fibrin
normal values 100-400mg/dl
Other: C3; beta- lipoprotein (LDL)

22
Receptor-mediated transferrin
endocsytosis
 In conditions associated
with increased need for
iron (iron deficiency or
late pregnancy) TIBC is
increased, but
saturation is decreased
from the normal 33%.
In hemochromatosis,
TIBC is low, but it is
saturated.
 Specific Proteins in Plasma (7)

Gamma-globulin
There are several classes; IgG,
IgA, IgM, IgD, IgE
All are comprised of
Two heavy chain (55Kd)
There are five types of heavy chain;
g, a, m, d, e
Two light chain
There are two type of light chain; k,
l
25
Some examples of different classes of
immunoglobulin
IgG
The major class in the body
Found in blood steam as well as tissue
IgA
secretary IgA = two IgA join by J-chain
secreated into mucus, intestinal juice
and tear to prevent infection
IgM
Composed of 5 immunoglobulin
molecule
Formed very early in infection and
activate complementary very easily

26
 Normal

Polyclonal gammopathy pattern

http://erl.pathology.iupui.edu/labmed/Generator.cfm?Image=SPE
27
 Normal

Monoclonalprotein present

http://erl.pathology.iupui.edu/labmed/Generator.cfm?Image=SPE
28