PLASMA PROTEIN

Obtained by centrifuging blood with

an anticoagulant
Contains fibrinogen + albumin and
globulins
SERUM
The fluid portion of the blood
obtained after removal of the fibrin
clot and blood cells
SERUM=PLASMA-FIBRINOGEN

CONTENTS OF PLASMA PROTEIN
0.9% inorganic ions, 0.8% small
organic molecules and 7% protein
Total protein content of N.plasma
6 -8 gm/dl
Albumin 3.5 -5 gm/dl
Globulins - 2.5 -3.5gm/dl
Fibrinogen 200-400mg/dl
Alb : glb ratio -1.2:1 to 1.5:1



CHARACTERICTIC OF PLASMA PROTEIN
Most of them are synthesized in the
liver Albumin and fibrinogen; 50 -
80% Globulin
Rest of the globulin Lymphoid tissue
Almost all are glycoproteins
Each one has a characteristic half-life
in the circulation






Plasma protein
Specific
plasma
protein
Albumin
Transport
protein
Acute phase
protein
Immunoglobulins
Complement
protein
Cytokines
Coagulation
factors
Plasma
enzymes




ALBUMIN
Most abundant plasma protein
Synthesized by hepatocytes used in
LFT
A low molecular weight protein
(69,000D)
Significant component of most ECF
Single polypeptide chain 585 amino
acids with 17 intra chain S-S bonds
Highly soluble in water
Half life 15-19 days

Role in Transport:
Transports metals, ions, fatty acids, amino
acids, bilirubin, enzymes, drugs etc..
Maintainance of colloidal osmotic
pressure
Buffering action


FUNCTIONS OF ALBUMIN
Maintenance of colloidal oncotic
pressure
Proteins exert effective osmotic
pressure 25mm Hg; 80%
contributed by Albumin
STARLINGS HYPOTHESIS :
At the capillary end;
BP / Hydrostatic pressure expels
water out
Effective osmotic pressure takes
water into the vascular compartment


CAUSE FOR EDEMA ?....
Arterial end of capillary BP -
35mm Hg
EOP - 25 mm Hg
Thus water expelled by a P of 10 mm Hg
Venous end - BP -
15mm Hg
EOP - 25 mm Hg
Thus water is imbibed with P of 10 mm Hg
Therefore blood volume remains the
same
If S.protein reduces;EOP decreases;
leads to accumulation of water

CAUSES OF HYPOALBUMINAEMIA
DECREASE IN ALBUMIN SYNTHESIS
Cirrhosis of liver disease (chronic
diseases)
Malnutrition
Haemodilution
a. Over hydration
b. Late stage of pregnancy
C. Artefactual-Blood drawn from driparm

INCREASED ALBUMIN SYNTHESIS
Nephrotic syndrome
Extensive burns
Malabsorption due to gastro-
intestinal disease
Protein-losing enteropathy
a. Ulceration of the bowel
b. Lymphatic obstruction

ALPHA ALBUMIN

1
is comprised :
1-antitrypsin


1- acid glycoprotein

Alpha fetoprotein



1- lipoprotein HDL
2 consist of:


Ceruloplasmin
2-macroglobulin
Haptoglobin- binds to
hemoglobin

Ceruloplasmin
2 globulin;contains 95% of total
copper found in serum
Single polypeptide chain
Transport protein for Cu; Acute phase
protein
Synthesized in liver
Copper is absorbed and transported
to liver but absence of hepatocellular
Ptype ATPase prevents incorporation
of Cu into Cp

Wilsons disease
Hepatolenticular degenaration
Autosomal recessive
Mutation in gene coding for copper
binding ATPase(excretion of Cu from
cells)
Decrease in Cp levels
Marked increase in tissue copper
liver,brain,kidney
Increased urinary Cu and tissue Cu

Menkes disease
X-linked inherited disorder
Dietary Cu absorbed but cannot be
transported to blood due to genetic
absence of an intracellular Cu binding
ATPase
Cu accumulates affects vascular and
connective tissues

Beta and gamma- globulins
Beta-globulin:
Haemopexin
Transferrin
C-reactive protein
2-macroglobulin
LDL lipoprotein

Gamma-globulin:
Immunoglobulins
Ig G, Ig M, Ig A, Ig D,Ig E
Transferrin
Principal plasma transport protein for
iron
Plasma levels regulated by levels of
iron
One molecule binds 2 molecules of
iron
Fe-Tf complex is taken up by all
tissues by specific surface receptors

Lipoproteins
Function to transport cholesterol, TAG
& phopholipid in blood
Sub classified into VLDL, IDL, LDL, HDL






Acute phase response and proteins
Nonspecific response to tissue injury
or infection Assessment of the
presence and degree of inflammation
Positive APPs: Increase in
inflammation
Negative APPs: Decrease in
inflammation
Presence of inflammatory disease
Monitoring therapeutic effectiveness
Follow-up of patient with malignancy

Acute phase reactants
Positive acute phase proteins
C-reactive protein
1-antitrypsin
1-antichymotrypsin
1-acid glycoprotein
Ceruloplasmin
Haptoglobin
Complement component C3 and C4
Negative acute phase proteins
Albumin
Transferrin
Pre-albumin








Estimation of serum proteins
Total protein estimation:
Biuret Method
Colorimetric method
Most widely used
Principle:
Alkaline Cu
2+
reacts with
compounds containing 2 or
more peptide bonds to give a
violet-colored complex, which
is then measured at 540 nm
Analysis of proteins
Specific quantitative assays-
immunochemical methods
nephelometry
turbidometry
electroimmunoassay
RIA, fluorescence immunoassay
chemiluminescence
Structural and quantitative information -
Mass specrometry

Estimation of s.proteins..
Estimation of serum albumin
Salt precipitation: Globulins
are precipitated in high salt
concentration; albumin in
supernatant is quantitated by
biuret reaction
Dye binding: Albumin binds to
dye; causes a shift in the
absorption maximum
BCG (bromocresol
green)
BCP (bromocresol
purple)

Electrophoresis
Technique invented by Tiselius in
1937,Nobel prize in 1948
Proteins separated based on electric
charge
The process of moving charged
molecules in solution by applying an
electrical field across the mixture
Molecules move with a speed
dependent on their charge, shape,
and size








PLASMA
PROTEIN
Total
serum
protein
Amount
in
grams
per
deciliter
(g/dL)
Amount in
SI units
grams per
liter (g/L)
Albumin 58%74% 3.55.5 3555
Alpha-1
globulin
2.0%
3.5%
0.20.4 24
Alpha-2
globulin
5.4%
10.6%
0.50.9 59
Beta
globulin
7%14% 0.61.1 611
Gamma
globulin
8%18% 0.71.7 717

Serum Total protein - 6.4 to 8.3 g/dL
Albumin - 3.5 to 5.0 g/dL
Serum globulin - 2.5 to 3.5 g/dL

Paraprotein
Is an abnormal plasma protein
appearing in large quantity; ELOMA
Malignant proliferation of
plasma cells; S. Ig G /Ig A Multiple myeloma
Light chains(low mol wt.) present in
urine are considered as Bence Jones
proteins
Starts precipitating on heating urine
at 45 - 60

and redissolves at above

80

C



Monoclonal gammopathy






MULTIPLE MYELOMA