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Plasma Proteins
GLOBULINS
By
N.Santhosh Kumar
Asst.Professor
Department of Biochemistry
SIMS & RH
• Globulins is a glycoprotein with mol wt – 90 to 1300kDa.
• There are separated by half saturation with ammonium sulphate.
• Main functions: Transport &Immunity.
• Globulins contributes different fractions in electrophoresis (α1,α2,β, γ
-globulins).
• Normal range of Globulins -1.8 to 3.6 gm%
Fraction Examples c (gm%)
1-globulins 1-antitrypsin, 1-acid glyco- protein, RBP,
1-lipoprotein (HDL), 1-fetoprotein, 0.1 to 0.4
2-globulins Haptoglobin, Ceruloplasmin
2-Macroglobulin, Thyroxin-binding
globulin (TBG),
0.4 to 0. 8
-globulins C-reactive protein (CRP) Transferrin,
Hemopexin, 2-Micro globulin, -lipoprotein
(LDL)
0.5 to 1.2
-globulins IgG, IgA, IgM, IgE, IgD, 0.7 to 1.5
α1 –antitrypsin (AAT) / α1-protease inhibitor
Glycoprotein(mol wt – 54kDa), synthesized by liver
AAT Inhibits activity of enzyme elastease – degrades elastin
protein (elasticity to the lungs)
AAT deficiency is an inherited disorder causes lung disease
(emphysema)& liver disease (neonatal hepatitis)
Emphysema (Abnormal distension of lungs by air)
Chronic obstructive lung disease, characterized by alveolar
damage
Enlarged alveoli
Compressed
alveoli duct
Collapsed
alveoli sac
Causes shortness of breath due to increasing of air sacs in the
alveolar, leading to difficulty in breathing
In chronic smoking
α1-feto globulin
• High conc. in foetal blood during mid pregnancy.
• Immunoregulatory during pregnancy
• Normal adult blood has less than 1 μg/100ml
• Increased in pregnancy, open neural tube defect (congenital
abnormalities caused by failure of the neural tube) Hepatoma
(cancer of liver)
• Decreased in Down’s syndrome (form of mental subnormality due
to a chromosome defect)
α2-macro globulin
 large plasma glycoprotein, synthesized by Monocytes, Hepatocytes &
macrophages with mol wt 7,20,000.
Functions:
 Carries of many growth factors such as platelet derived growth factor.
 Inactivates all the proteases – invivo Anticoagulant.
 Natural inhibitor of endopeptidase such as trypsin, plasmin, chymotrypsin
& thrombin etc.
Clinical significance:
Normal serum level – 130 – 300 mg/dl.
 Increased levels - Nephritic syndrome, cirrhosis
Collagen disorders
 Decreased levels - myeloma, Pancreatitis, Peptic ulcer
Haptoglobin (α2 globulin)
 syn in liver with half life is 5days.
 It binds and clears the free Hb found outside of RBCs from circulation.
Hb-Hp half life is 90min.
 Helps in the breakdown of Hb to bilirubin.
 Prevents the loss of iron in the urine
Clinical significance :
Levels in serum 40-170mg/dl
 Increased in Inflammatory conditions
Rheumatic diseases.
 Decreased: intravascular haemolytic anemia
- Hp assay is used to screen & monitor intravascular haemolytic
anaemia
Ceruloplasmin
 It is a blue colored α2-globulin, syn in liver .
 Major transport protein for copper (6 - 8 cu atoms /mol).
 Shows ferroxidase activity (helps in oxidation of Fe2+ to Fe3+.
Clinical significance :
Normal levels – 25 -50 mg/dl
Increased levels
Inflammatory conditions
Collagen disorders
Decreased levels
Wilson's disease,
Malnutrition
Nephrotic syndrome
C - reactive proteins
 It is a β-fraction of globulin with 115-140 KD & binds
to T- lymphocytes.
 CRP was discovered in serum of pneumococcal
pneumonia patients.
 CRP reacts with C-polysaccharides of Capsule of
pneumococci bacteria.
 Stimulated complement activity, Phagocytosis by
macrophages - clears necrotic cells , apoptotic cells
and bacteria.
 Useful in differentiating bacterial from viral infections. because CRP levels
increased in bacterial infections only
γ- globulins
 γ- fraction of globulins are
immunoglobulin / antibodies.
 Produced by plasma cells
 Igs provide resistance because
they can neutralize toxins &
viruses, Opsonize microbes,
Activate complement & prevent
the attachment of microbes to
mucosal surfaces
Types and functions of Immunoglobulins
Igs Structure /
Characteristics
Found in Total %
in Ig
Functions
IgG Exists as simple Y shaped
structure. Can cross the
placental barrier.
Blood,
lymph fluid
& CSF
85% -Involved in precipitation, complement
fixation reactions & neutralization of
toxin compounds.
IgA Exists as dimer joined by a
J chain
Saliva,
tears, sweat
10-13% - Protects the body surface from foreign
substance.
IgM Largest antibody exists as
pentamer held together by J
chain. Primary antibody
and acts as first line
defense
Body fluids 5-10% - Protects against invading micro-
organisms.
- Promotes the phagocytosis,
agglutination, bacteriolysis reactions.
Igs Structure / Characteristics Found in Total %
in Ig
Functions
IgM Largest antibody exists as
pentamer held together by J
chain. Primary antibody and
acts as first line defense
Body fluids 5-10% - Protects against invading micro-
organisms.
- Promotes the phagocytosis,
agglutination, bacteriolysis reactions.
IgE Exists as simple Y shaped
structure
Lungs, skin,
& mucous
membrane
5% - Receptor for allergens & parasitic
antigens.
- Protection against parasitic infections.
- Responsible for anaphylactic
hypersensitivity.
IgD Exists as simple Y shaped
structure
Body fluids 1% Serve as recognition of receptors for
antigen.
Acute phase proteins
&
Acute phase reactants (APRs)
Def:
levels of certain proteins in plasma increases (or) decreases
during acute inflammatory states (or) secondary to certain types of
tissue damage.
 APRs concentration changes in:
 infection
 surgery
 injury
 cancer
acute inflammatory
states
Mechanism of AFPs
TYPES OF ACUTE PHASE PROTEINS
Positive A.P.P
• 1-antitrypsin
• α2- macroglobulin
• C-reactive protein (CRP)
• Haptoglobin (HP)
• Ceruloplasmin
Liver disease, trauma, burns
Nephritic syndrome, collagen
disorders
Inflammatory conditions,
malignancies, collagen disorders
Hepatocellular damage
&nephrotic syndrome.
Myocardial infarction,
inflammations, trauma
&stress
Negative A.P.P
• Albumin
• Transthyretin/TBP
•Transferrin
Inflammation, liver diseases,
nephrotic syndrome
malnutrition
Inflammation, malignancy,
nephrotic syndrome
Liver diseases, thyrotoxicosis,
malnutrition
TYPES OFACUTE PHASE PROTEINS
TRANSPORT PROTEINS
Transport Proteins Transports
Pre albumin / transthyretin
(Thyroxine binding prealbumin
Thyroid hormones of thyroxin,
Retinol, steroid hormones
Albumin Bilirubin, Free fatty acids, Calcium,
Drugs etc
Retinol binding protein Vitamin - A (retinol).
Thyroid binding globulin Thyroid hormones
Haptoglobin Hemoglobin
Transferrin Iron (Ferrous state)
HDL Carries cholesterol from peripheral
tissues to liver
LDL Carries cholesterol from liver to
peripheral tissues.
Case base Question
Patient gives history of cough, fever since 1 month and dyspnea
(difficulty in breathing) since 4 days. Family history reveals that
one of his brother had died earlier due to lung disease
Q a) What could be the probable diagnosis?
b) What is the biochemical defect?
c) Explain this with respect to chronic smoking?
Separation Of Plasma Proteins
Electrophoresis
Based on the migration of charged solutes of a solution towards the opposite
electrical poles in an electric field.
Multiple myeloma
M-band between beta and gamma region.
Q) a) What is the probable diagnosis?
b) Mention the biochemical defect
Chronic infection,
broad based increase in gamma region; general increase in alpha-1 and alpha-2
bands; decreased albumin band
Nephrotic syndrome
hypoalbuminemia; prominent alpha-2 band
Cirrhosis of liver;
γ-β- bridge observed (increased γ & β- globulins) &
decreased albumin band
Agammaglobulinemia ;
Absent or decreased γ globulins observed
& other bands are normal
Q) a) What is the probable diagnosis?
b) Mention the biochemical defect.
Next PPT on
PP-03: Gamma globulins (Immunoglobulins) &
Multiple Myeloma
N.Santhosh Kumar /Asst.Professor of Biochem / SIMSRH
Thank you
The End

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PP-02: Plasma Proteins (Globulins, Acute phase proteins, Transport proteins)

  • 2. • Globulins is a glycoprotein with mol wt – 90 to 1300kDa. • There are separated by half saturation with ammonium sulphate. • Main functions: Transport &Immunity. • Globulins contributes different fractions in electrophoresis (α1,α2,β, γ -globulins). • Normal range of Globulins -1.8 to 3.6 gm%
  • 3. Fraction Examples c (gm%) 1-globulins 1-antitrypsin, 1-acid glyco- protein, RBP, 1-lipoprotein (HDL), 1-fetoprotein, 0.1 to 0.4 2-globulins Haptoglobin, Ceruloplasmin 2-Macroglobulin, Thyroxin-binding globulin (TBG), 0.4 to 0. 8 -globulins C-reactive protein (CRP) Transferrin, Hemopexin, 2-Micro globulin, -lipoprotein (LDL) 0.5 to 1.2 -globulins IgG, IgA, IgM, IgE, IgD, 0.7 to 1.5
  • 4. α1 –antitrypsin (AAT) / α1-protease inhibitor Glycoprotein(mol wt – 54kDa), synthesized by liver
  • 5. AAT Inhibits activity of enzyme elastease – degrades elastin protein (elasticity to the lungs) AAT deficiency is an inherited disorder causes lung disease (emphysema)& liver disease (neonatal hepatitis)
  • 6. Emphysema (Abnormal distension of lungs by air) Chronic obstructive lung disease, characterized by alveolar damage
  • 7. Enlarged alveoli Compressed alveoli duct Collapsed alveoli sac Causes shortness of breath due to increasing of air sacs in the alveolar, leading to difficulty in breathing
  • 9. α1-feto globulin • High conc. in foetal blood during mid pregnancy. • Immunoregulatory during pregnancy • Normal adult blood has less than 1 μg/100ml • Increased in pregnancy, open neural tube defect (congenital abnormalities caused by failure of the neural tube) Hepatoma (cancer of liver) • Decreased in Down’s syndrome (form of mental subnormality due to a chromosome defect)
  • 10. α2-macro globulin  large plasma glycoprotein, synthesized by Monocytes, Hepatocytes & macrophages with mol wt 7,20,000. Functions:  Carries of many growth factors such as platelet derived growth factor.  Inactivates all the proteases – invivo Anticoagulant.  Natural inhibitor of endopeptidase such as trypsin, plasmin, chymotrypsin & thrombin etc.
  • 11. Clinical significance: Normal serum level – 130 – 300 mg/dl.  Increased levels - Nephritic syndrome, cirrhosis Collagen disorders  Decreased levels - myeloma, Pancreatitis, Peptic ulcer
  • 12. Haptoglobin (α2 globulin)  syn in liver with half life is 5days.  It binds and clears the free Hb found outside of RBCs from circulation. Hb-Hp half life is 90min.  Helps in the breakdown of Hb to bilirubin.  Prevents the loss of iron in the urine
  • 13. Clinical significance : Levels in serum 40-170mg/dl  Increased in Inflammatory conditions Rheumatic diseases.  Decreased: intravascular haemolytic anemia - Hp assay is used to screen & monitor intravascular haemolytic anaemia
  • 14. Ceruloplasmin  It is a blue colored α2-globulin, syn in liver .  Major transport protein for copper (6 - 8 cu atoms /mol).  Shows ferroxidase activity (helps in oxidation of Fe2+ to Fe3+. Clinical significance : Normal levels – 25 -50 mg/dl
  • 15. Increased levels Inflammatory conditions Collagen disorders Decreased levels Wilson's disease, Malnutrition Nephrotic syndrome
  • 16. C - reactive proteins  It is a β-fraction of globulin with 115-140 KD & binds to T- lymphocytes.  CRP was discovered in serum of pneumococcal pneumonia patients.  CRP reacts with C-polysaccharides of Capsule of pneumococci bacteria.  Stimulated complement activity, Phagocytosis by macrophages - clears necrotic cells , apoptotic cells and bacteria.
  • 17.  Useful in differentiating bacterial from viral infections. because CRP levels increased in bacterial infections only
  • 18. γ- globulins  γ- fraction of globulins are immunoglobulin / antibodies.  Produced by plasma cells  Igs provide resistance because they can neutralize toxins & viruses, Opsonize microbes, Activate complement & prevent the attachment of microbes to mucosal surfaces
  • 19. Types and functions of Immunoglobulins Igs Structure / Characteristics Found in Total % in Ig Functions IgG Exists as simple Y shaped structure. Can cross the placental barrier. Blood, lymph fluid & CSF 85% -Involved in precipitation, complement fixation reactions & neutralization of toxin compounds. IgA Exists as dimer joined by a J chain Saliva, tears, sweat 10-13% - Protects the body surface from foreign substance. IgM Largest antibody exists as pentamer held together by J chain. Primary antibody and acts as first line defense Body fluids 5-10% - Protects against invading micro- organisms. - Promotes the phagocytosis, agglutination, bacteriolysis reactions.
  • 20. Igs Structure / Characteristics Found in Total % in Ig Functions IgM Largest antibody exists as pentamer held together by J chain. Primary antibody and acts as first line defense Body fluids 5-10% - Protects against invading micro- organisms. - Promotes the phagocytosis, agglutination, bacteriolysis reactions. IgE Exists as simple Y shaped structure Lungs, skin, & mucous membrane 5% - Receptor for allergens & parasitic antigens. - Protection against parasitic infections. - Responsible for anaphylactic hypersensitivity. IgD Exists as simple Y shaped structure Body fluids 1% Serve as recognition of receptors for antigen.
  • 21. Acute phase proteins & Acute phase reactants (APRs)
  • 22. Def: levels of certain proteins in plasma increases (or) decreases during acute inflammatory states (or) secondary to certain types of tissue damage.  APRs concentration changes in:  infection  surgery  injury  cancer
  • 25. TYPES OF ACUTE PHASE PROTEINS Positive A.P.P • 1-antitrypsin • α2- macroglobulin • C-reactive protein (CRP) • Haptoglobin (HP) • Ceruloplasmin Liver disease, trauma, burns Nephritic syndrome, collagen disorders Inflammatory conditions, malignancies, collagen disorders Hepatocellular damage &nephrotic syndrome. Myocardial infarction, inflammations, trauma &stress
  • 26. Negative A.P.P • Albumin • Transthyretin/TBP •Transferrin Inflammation, liver diseases, nephrotic syndrome malnutrition Inflammation, malignancy, nephrotic syndrome Liver diseases, thyrotoxicosis, malnutrition TYPES OFACUTE PHASE PROTEINS
  • 28. Transport Proteins Transports Pre albumin / transthyretin (Thyroxine binding prealbumin Thyroid hormones of thyroxin, Retinol, steroid hormones Albumin Bilirubin, Free fatty acids, Calcium, Drugs etc Retinol binding protein Vitamin - A (retinol). Thyroid binding globulin Thyroid hormones Haptoglobin Hemoglobin Transferrin Iron (Ferrous state) HDL Carries cholesterol from peripheral tissues to liver LDL Carries cholesterol from liver to peripheral tissues.
  • 29. Case base Question Patient gives history of cough, fever since 1 month and dyspnea (difficulty in breathing) since 4 days. Family history reveals that one of his brother had died earlier due to lung disease Q a) What could be the probable diagnosis? b) What is the biochemical defect? c) Explain this with respect to chronic smoking?
  • 30. Separation Of Plasma Proteins Electrophoresis Based on the migration of charged solutes of a solution towards the opposite electrical poles in an electric field.
  • 31. Multiple myeloma M-band between beta and gamma region. Q) a) What is the probable diagnosis? b) Mention the biochemical defect
  • 32. Chronic infection, broad based increase in gamma region; general increase in alpha-1 and alpha-2 bands; decreased albumin band
  • 34. Cirrhosis of liver; γ-β- bridge observed (increased γ & β- globulins) & decreased albumin band
  • 35. Agammaglobulinemia ; Absent or decreased γ globulins observed & other bands are normal
  • 36. Q) a) What is the probable diagnosis? b) Mention the biochemical defect.
  • 37. Next PPT on PP-03: Gamma globulins (Immunoglobulins) & Multiple Myeloma N.Santhosh Kumar /Asst.Professor of Biochem / SIMSRH