Professional Documents
Culture Documents
Prof.Dr.Gülden Burçak
2020-2021
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Immunoglobulins
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Immunoglobulins
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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
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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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