Immunoglobulins

Prof.Dr.Gülden Burçak
2020-2021

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 Immunoglobulins

• Elements of humoral immune system

• Proteins produced by the plasma cells of the bone marrow as
part of the immune response
• The plasma cells are B lymphocytes transformed after
exposure to a foreign (or occasionally an endogenous) antigen
• Adaptive immune response
 Specificity
 Memory
• Glycoproteins in serum and tissue fluids, some are carried on
B-lymphocytes

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 Immunoglobulins

• Five distinct classes

• IgG, IgA, IgM, IgD and IgE
• Differ in amino acid composition, carbohydrate content,
charge and size
• Heterogeneity within each class
• α- γ electrophoretic mobility
 α in Ig A, γ in IgG, μ in IgM, δ in IgD and ε in IgE

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Plasma protein electrophoresis pattern in agarose gel is composed
of five fractions, each composed of many individual species
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 Two identical heavy chains and two identical light chains
 Heavy chains, MW 50000 -77000
 Structurally distinct for each class and subclass (IgG and IgA
have subclasses)
 Light chains, MW 25 000, two types (κ or λ)
 Any type of light and heavy chain combination
 Linked by covalent (disulfide bridges) and non-covalent bonds
(intra-, inter-chain)

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Bifunctional molecules
1) Fab : Binding to the antigen with variable and hypervariable
amino acid sequences
2) Fc : crystallizable fragment which has effector function,
binding to
 host tissues
 various cells of the immune system
 some phagocytic cells
 first component of the classical complement system (C1q)

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Humans are capable of producing more than 108 different
antibodies with distinct binding specificities
Antibody diversity is derived from random reassembly of a
set of immunoglobulin gene segments through genetic
recombination mechanisms
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• Specificity is conferred by chemical complementarity
between the antigen and its specific binding site, in terms of
shape and the location of charged, nonpolar, and hydrogen-
bonding groups
• Complementarity is mostly achieved interactively as the
structures of antigen and binding site influence each other as
they come closer together
• Conformational changes in the antibody and/or the antigen
then allow the complementary groups to interact fully
• Flexibility in the hinge region : rich in glycine

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 Immunoglobulin G
• Major Ig in serum
• 70-75 % of total Ig pool, distributed evenly between
intravascular and extravascular pools
• A single four chain molecule
• Four subclasses (γ1, γ2, γ3, γ4)
• Major antibody of secondary immune response
• Exclusive antitoxin class
• Maternal IgG confers immunity in neonates
• Activates complement and chemotactically attractsPMN
leukocytes

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The basic
structure of IgG1

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 Immunoglobulin A
• 15-20 % of human serum Ig pool
• Two subclasses α1 and α2
• In man 80 % of IgA is monomeric IgA1
• Secretory IgA (sIgA) is mainly IgA2
 Two units of Ig A, one J-chain,one secretory component
( protects against proteases )
 predominant Ig in seromucous secretions ( saliva, colostrum,
milk, tracheobronchial and genitourinary secretions )
 intestine, mammary ducts

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 Immunoglobulin M
• 10 % of total Ig pool
• A pentamer of the basic four chain structure, J –chain
• Largely confined to intravascular pool
• Predominant early antibody
• The most efficient activator of complement
• Ig molecule synthesized in the newborn

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 Immunoglobulin D
• < 1% of total plasma Ig
• Present in large quantities on the membrane of many B-cells
• Precise biologic function is unknown
• Susceptible to heat and proteases

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 Immunoglobulin E
• Scarce in serum
• On surface membrane of basophils and mast cells
• Sensitizes cells on mucosal surfaces ( conjunctival, nasal and
bronchial mucosa )
• Immunity to helminithic parasites
• Commonly associated with allergic diseases
 asthma and hay fever

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 Increase in Immunoglobulins

• Igs may be increased non-specifically in a wide variety of

infections and also in autoimmune diseases
• If increased synthesis comes from a number of cell lines, each
producing its own specific Ig, a diffuse increase in protein
mass is observed on the γ globulin region on electrophoresis
• Polyclonal Igs are slightly different from each other in size and
charge, dont not migrate to the same place on electrophoresis
• Polyclonal hypergammaglobulinemia can result from chronic
inflammation or other disorders such as cirrhosis of the liver

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• Monoclonal Igs synthesized from a single clone, are identical
and observed on electrophoresis as a single discrete band
• An intact monoclonal Ig or a fragment and is called a
paraprotein
• Multiple myeloma : Malignant neoplasm of a single clone of
plasma cells of the bone marrow
 Bence-Jones protein : Light chains of Ig molecules
 precipitates between 45-60°C, dissolves on further heating
but reappears when the solution is cooled to about 60°C
 found in 35-65% of the patients

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• Waldenström’s macroglobulinemia : A single monoclonal IgM
causes a hyperviscosity syndrome, very thick consistency of
the circulating blood that can be exacerbated by cold ambient
temperatures
• Cryoglobulinemia : Presence of a serum protein which
precipitates below 37°C
• Cryoglobulins are Igs that precipitate upon storage at 4° C, and
associated with hematologic, autoimmune, chronic infections
such as hepatitis C

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• Amyloidosis : Extracellular deposit of amyloid consisting an
insoluble fibrillar protein (β-conformation) and a glycoprotein
resembling starch and resistant to proteolysis
 from misfolded Ig light chains or fragments of light chains
• Autoimmune disease : Formation of antibodies against
native antigens.
• Hypoglobulinemia or agammaglobulinemia is often the prime
characteristic of some immunodeficiency disorders
• Deficiencies or absence of Igs can occur as a result of
infection, genetic abnormalities or the effects of therapy
 treatment with Ig-rich plasma or the transplantation of bone-
marrow-containing competent plasma cells
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Enzyme-Linked Immunosorbent Assay ( ELISA )
• The extraordinary binding affinity and specificity of monoclonal
antibodies make them valuable analytical reagents
• Detection and quantification of an antigen (protein) in a sample
• Proteins in the sample are adsorbed to an inert surface
• Unoccupied sites are coated with a nonspecific protein
• The surface is then treated with the primary antibody ( Ab )
• After incubation, the unbound Ab is washed off and a secondary
Ab, against the primary Ab linked to an enzyme, is added
• The substrate of the enzyme is added, a colored product forms
 the formation of the color indicates the presence of the protein
in the sample
 the intensity of the color measured gives the concn of it
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