Why is it important for our body to have plasma proteins in our

blood?

How do plasma proteins affect our health?

Case Based Scenario
A 60-year-old female. • Examination findings:
Pitting edema (swelling that leaves an
• Presenting Complaint: Swelling in her legs indentation) in both legs and around her
and face for the past 3 weeks. She also eyes.
feels very tired and has had a poor
• Blood Tests: Serum Albumin: 2.0 g/dL
appetite recently.
(Normal: 3.5–5.0 g/dL)
Serum Creatinine: 1.0 mg/dL (Normal)
• History: Mrs. Verma has a h/o high blood
pressure and diabetes for 15 years. She Blood sugar: 170 mg/dL (High, indicating
has been taking medications for both, but poor blood sugar control)
hasn't been very regular with her diet or • Urine Test:
medications. She is not known to have Protein in urine: 2+ (Proteinuria, which
any kidney or liver diseases. indicates a lot of protein in the urine)
 Dr. Shaheena Yassir
Associate Professor
Department of Biochemistry
3/12/2024

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 SPECIFIC LEARNING OBJECTIVES

• To define plasma and plasma protein

• To identify the major plasma proteins and their synthesis
• To enumerate normal levels of plasma proteins
• To brief different ways of separation of plasma proteins
• To illustrate normal and abnormal electrophoretic pattern of
serum protein
• To explain functions and clinical significance of albumin
• To brief subclasses of globulin with examples 4
• Blood = plasma + cells
(RBC+WBC + platelets).
• Plasma = blood – cells
• Serum = plasma – clot
(fibrinogen and other
clotting factors)

Serum separates after standing

blood for 15 – 20 minutes.

5
 Plasma proteins

Plasma proteins are the proteins found in blood plasma, the pale-yellow
liquid portion of blood. They are primarily synthesized by the liver, and
they are essential for various bodily functions.
Albumin : The most abundant protein
Globulins: Globulins are classified based on electrophoretic
separation
α− globulin β− globulin γ− globulins
Fibrinogen 6
 Synthesis of Plasma Proteins

• All the albumin and fibrinogen are synthesized by the liver only
• Similarly 50 to 80% of the globulin is formed in the liver
• The remainder of the globulins are formed almost entirely in the
lymphoid tissues.
• The A/G ratio can be altered in the liver disease.

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 Plasma Protein - Normal values

The normal serum protein level is 6.0-8.0 g/ dL.

a. Albumin – 3.5 - 5.5g/dL (the most abundant protein
b. Globulins: 2.5 - 3.5g/dL
(Globulins are classified based on electrophoretic separation)
• α− globulin: α1 & α2−globulins
• β− globulin: β1 & β2 -globulins
• γ− globulins
c. Fibrinogen 200 to 400 mg%
8
 Separation of Plasma Proteins

The plasma proteins can be separated by :

- Salting out
- Electrophoresis
- Ultracentrifugation
- Immunoelectrophoresis.

9
 Separation of Plasma Proteins

[Link]
10
[Link]
Electrophoretic pattern of normal serum
protein

Albumin - 55 - 65%
Alpha 1 globulin: 2 - 4%
Alpha 2 globulin: 6 -12%
Beta globulin: 8 - 12%
Gamma globulin: 12 - 22%

[Link]

12
Abnormal patterns in clinical disease

13
[Link]
•[Link].69000, 585 amino acids.

•Synthesised exclusively in liver

•Half-life – 20 days

•Serum level – 3.5 to 5.4 gm/dL

[Link]
mics_Simulation_and_Free_Energy_Studies_on_the_Interaction_of_Sali
cylic_Acid_with_Human_Serum_Albumin_HSA/figures?lo=1

14
 Functions of albumin
1. To maintain colloidal osmotic pressure

[Link]

15
[Link]

16
 Functions of albumin

2. Transport of hydrophobic substances (bilirubin, free fatty acids),

metal ions (Ca++, Cu++, Zn++) and hormone, thyroxine.

3. Buffering: The histidyl residues in all plasma proteins act as a

buffer. Plasma proteins, including, albumin are the second most
important buffer after bicarbonate buffer in the blood.

4. Supply of amino acid to cells: Cells take up albumin by pinocytosis

and degrade it, thus getting a supply of essential and other amino
17
acids from the liver.
Clinical significance

• Hypoalbuminemia (most common)

• Hyperalbuminemia
Causes of Hypoalbuminemia (<3.5 g/dL)

1. Decreased Synthesis

2. Increased Loss

3. Redistribution or Dilution
1. Decreased Synthesis

Liver Disease:

• Chronic liver diseases (e.g., cirrhosis, hepatitis)

• Acute liver failure
Malnutrition:

• Protein-calorie malnutrition
• Severe malabsorption (e.g., celiac disease, Crohn’s disease)
Chronic Illness
2. Increased Loss
Kidney Diseases:
• Nephrotic syndrome (protein loss in urine)
• Chronic kidney disease
Gastrointestinal Loss:
• Protein-losing enteropathies (e.g., Crohn's disease, intestinal lymphangiectasia)
Skin Loss:
• Severe burns
• Exudative skin conditions (e.g., toxic epidermal necrolysis)
Blood Loss:
• Chronic or acute hemorrhage
3. Redistribution or Dilution

Increased Vascular Permeability:

• Sepsis or systemic inflammation (e.g., due to infections, trauma)

• Capillary leak syndrome
Fluid Overload:

• Excessive IV fluids
• Congestive heart failure (leading to dilutional hypoalbuminemia)
 • Clinical manifestation: decrease of
colloidal osmotic pressure of blood which
causes reduced fluid re-entry from ECF into
the capillaries and therefore edema.
• Biochemical Investigations:
[Link]

• a) Estimation of serum albumin and

• b) Electrophoresis of serum proteins
23
 Clinical significance

Hyperalbuminaemia: Acute dehydration have no clinical significance.

Analbuminaemia (less than 1gm/l , normal= 3.5 to 6gm/dl):

Analbuminaemia is a rare hereditary abnormalities there may be no

symptoms or signs mild edema due to compensatory increase in

plasma globulin concentration.

 GLOBULINS:

• 35% of plasma proteins are

globulins
• They are of two types
immunoglobulins and
transport globulins

[Link]

25
 Subclassification of globulins
Alpha 1 globulin

• Retinol binding protein - Transport of vitamin A

• Alpha fetoprotein - Embryonic protein
• Alpha 1 antitrypsin. - Antiprotease (inhibitor of elastase
• Apha 1 acid glycoprotein - Tissue repair

26
• It’s a glycoprotein, a serine protease inhibitor

• It combines with trypsin, chymotrypsin, elastase,

cathepsin, plasmin and thrombin, and inhibits their activity.

• Its deficiency is implicated in emphysema and cirrhosis of

the liver.

• It protects plasma & tissue proteins by inhibiting

proteases , it combines with elastase, trypsin and protects
the hydrolytic damage of tissues such as lungs.
Alpha 2 globulin
• Haptoglobulin - Binds to free haemoglobin
• Ceruloplasmin - Transport of copper
• Alpha 2 macroglobulin - Natural antiprotease inhibits thrombin,
trypsin and pepsin

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 Haptoglobin
Plasma glycoprotein with 90,000 molecular
weight .
FUNCTIONS

[Link] to free Hb and prevents its loss via

kidney
CLINICAL SIGNIFICANCE

levels decrease in hemolytic anaemia

Ceruloplasmin

• α2 globulin ,synthesized in liver, acute phase protein

• 6-8 copper atoms per molecule

• It helps in the incorporation of iron into transferrin.

• It is an important antioxidant

• Ceruloplasmin deficiency - Wilson’s hepatolenticular degeneration

leading to cirrhosis due to accumulation of copper in liver, kidney,
brain, around the cornea and RBCs.
 α2Macroglobulin:

• It is a tetrameric protein

• synthesized by hepatocytes and macrophages.

• It inactivates all proteases and thus it is an important in vivo

anticoagulant by inhibiting thrombin.

• Its concentration is markedly increased in nephrotic syndrome.

Beta globulin
• Hemopexin - Binds heme and prevents loss of heme
• Transferrin - Transport of iron
• C reactive protein - Body defence mechanism
• Beta 2 microglobulin - Body defence mechanism

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Transferrin:

• Each transferrin molecule binds 2 Fe3+ and

transports it
• Transferrin also acts as an antioxidant (prevents
ROS)
• Transferrin levels are decreased in:
– liver disease (e.g. cirrhosis)
– Chronic infections
– Nephrosis
• Increased levels occur during increased transferrin
synthesis caused as a result of iron deficiency
anemia
FIBRINOGEN:
• 4% of plasma proteins,
• Function - blood clotting during which it is
converted to fibrin
• Many factors ( plasma proteins ) are
involved in clotting mechanism thus
preventing loss of excessive amount of
blood. [Link], VIII, thrombin etc.
• Deficiency leads to e.g hemophilia,
Anticoagulant activity (thrombolysis):
• Plasmin breaks down thrombin and
dissolves the clot
 Functions of plasma proteins
Functional Plasma proteins
Colloidal osmotic pressure Albumin
Immunoproteins γ-globulin (antibody), CRP
Clotting Fibrinogen and other factors
Transport Transferrin
Thyroxine binding prealbumin (TBPA)
Retinol binding protein (RBP)
Thyroxine binding globulin (TBG).
Haptoglobin
Hemopexin
Lipoproteins
α-1 acid glycoprotein 35
Functional Class Plasma proteins

Enzymes Ceruloplasmin
Enzymes Inhibitors α-1-antitrypsin
((protease inhibitors) α- 2 -macroglobulin.

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 Transport Proteins and their Functions

Transport Proteins Substance Transported

• Albumin - Bilirubin, Free FA, metal ions,

Thyroxine, steroid hormones
• Transferrin - Iron
• Thyroxine binding prealbumin (TBPA) - T3 and T4
• Retinol binding protein (RBP) - Retinol
• Transcortin - cortisol and corticosterone
• Thyroxine binding globulin (TBG) - T3 and T4
• Haptoglobin - Hemoglobin
• Hemopexin - Heme
• Lipoproteins - Triacylglycerol and Cholesterol
• α-1 acid glycoprotein - Some lipid soluble substances
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Acute phase proteins

–Positive acute phase proteins: Increase

during inflammation.
–Negative acute phase proteins: decrease
during inflammation.
• Play a defensive role during surgery, MI,
infections and tumours.
Positive: Negative:
C-reactive protein: albumin
~1000-fold increase!
transferrin
1-antitrypsin antithrombin
fibrinogen transcortin
haptoglobin (HP) retinol binding
2 macroglobulin protein
C-Reactive protein (CRP):

• Its named because it reacts with C-

polysaccharide of capsule of
pneumococci.
• It is a β globulin synthesized in liver.
• CRP level has a positive correlation
in predicting the risk of coronary
artery disease.
• Elevated levels of CRP are indicative
of chronic, low-grade inflammation .
 References
• D.M. Vasudevan. Text book of Biochemistry 9th
Edition.
• U. Satyanarayanana. Biochemistry
• Textbook of Biochemistry, Third Edition - Pankaja
Naik

41
Small group teaching

Immunoglobulin
Acute phase reactants

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 Immunoglobulins (Ig)

• The immunoglobulins are γ -globulins, called antibodies.

• Constitute about 20% of all the plasma proteins

• produced by plasma cells and by lymphocytes in response to a variety of

antigen.
Figure 27.3: Schematic structure of immunoglobulin G.

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▪ The basic immunoglobulin is Y shaped consist of four
polypeptide chains:
– two H and
– two L chains
▪ The four chains are linked by disulfide bond
▪ L chain may be either of two types, Kappa (қ) or Lambda (λ)
but not both
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▪ The heavy chains may be of five types and are designated by
Greek letter:
– alpha (α)
– gamma (γ),
– delta (δ),
– mu (μ) and
– epsilon (ε)
▪ Immunoglobulins are named as per their heavy chain type as
IgA, IgG, IgD, IgM and IgE 46
47
 Light chain type

• Light chain may be either of two types, Kappa (k) or

Lambda (l) but not both.

• In a given immunoglobulin either 2k or 2 l but not the

mixture of kappa and lambda

• Most abundant light chain in human is k

▪ Enzyme (papain) digestion splits the immunoglobulin molecule into two
fragments

1. Fab: Fragment for antigen binding. Located in variable region.

2. Fc: Crystallisable fragment or fragment for

complement binding
Monomeric and dimeric forms of immunoglobulins.
 References
• D.M. Vasudevan. Text book of Biochemistry 9th
Edition.
• U. Satyanarayanana. Biochemistry

54
Pentameric form of immunoglobulin
▪ Most common Ig in serum: IgG

▪ Least common Ig in serum: IgE

▪ Largest Ig: IgM

▪ Pentamer Ig: IgM

▪ Dimmer: Secretary IgA

▪ Ig fixes the complement: IgG and IgM

▪ Ig present in secretion: IgA

▪ Ig can cross placenta: IgG

▪ IgG can not cross placenta: IgG2Ig involved in primary

immune response: IgM

▪ Ig secondary immune response: IgG

▪ Ig with J chain: IgM and IgA

▪ Ig with secretary piece: IgA

 Disorders due to changes in immunoglobulins

Quantitative changes in the amount of immunoglobulins in the

plasma and urine are known in several pathological conditions in

human.
– Multiple Myeloma
– Amyloidosis
– Bence Jones Proteins
– Cryoglobulinaemia
– Waldenstrom’s Macroglobulinaemia
 References
• D.M. Vasudevan. Text book of Biochemistry 9th
Edition.
• U. Satyanarayanana. Biochemistry

59
 Assignments

[Link] the plasma proteins and their normal values.

[Link] the cause of hypoalbuminemia and consequences of

hypoalbuminemia.

[Link] the principle of electrophoresis. Explain the normal

electrophoretic pattern of serum proteins.

[Link] the immunoglobulins. Describe their functions.

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 Question Bank

5. Name and explain the functions of plasma proteins with clinical

significance

6. Bence Jones proteins and tests to detect Bence Jones proteins

7. Reverse A: G ratio

8. Acute phase reactants

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62