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Rabiul Haque
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Learning Objectives (Antibodies Part 1)
• Definition of Antibody
• Structure of Immunoglobulins
• Classes of Antibody
• Important Functions of Antibody
• Polyclonal & Monoclonal Antibodies
• Concept of Isotypes, Allotypes & Idiotypes
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Learning Objectives (Antibodies Part 2)
• Immunoglobulin Genes
• Immunoglobulin Class Switching
• Hybridoma Cells
• Production of Monoclonal Antibodies
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Definition of Antibodies
• Ref. Lange Review of Medical Microbiology & and Immunology 14th Edition
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Antibodies Are Gamma Globulins
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Structure of Immunoglobulin
• Immunoglobulins are glycoproteins and consist of light and
heavy polypeptide chains.
• Light chains (L) have a molecular weight of about 25,000.
• Heavy chains (H) have a molecular weight of 50,000 to
70,000.
• The simplest antibody molecule consists of four polypeptide
chains and are Y shaped.
• The four polypeptide chains include two heavy chains and two
light chains. (molecular formula H2L2)
• An individual antibody always contains identical heavy chains
and identical light chains.
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Structure of Immunoglobulin
• The light and heavy chains are subdivided into variable and
constant regions.
• The regions are made of three-dimensionally folded,
repeating segments called domains.
• Each domain is approximately 110 amino acids long.
• A light chain contains one variable domain and one constant
domain.
• Most heavy chains contain one variable and three constant
domains.
– IgG, IgD and IgA have 3 constant domains in their heavy chains.
– IgM and IgE have 4 constant domains in their heavy chains
making them longer and heavier.
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Structure of Immunoglobulin
Amino terminal end
VL
CL
VH
CH1
CH2
Interchain disulfide
bonds
CH3
CL
VH
Heavy chain
CH1
CH2
Interchain disulfide
bonds
CH3
CH2
Interchain disulfide
bonds
CH3
CH2
Constant domains of heavy chain
CH3
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Structure of Immunoglobulin
• Hypervariable Region:
– Both heavy and light chains have three extremely variable
amino acid sequences at the amino terminal end of their
variable regions that form antigen-binding sites.
– Five to ten amino acids in each hypervariable region form the
antigen -binding site.
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Structure of Immunoglobulin
Amino terminal end
VL
CL
VH Hypervariable regions
CH1 of heavy chain
CH2
Interchain disulfide
bonds
CH3
• Light chains
– Two types: (according to amino acid differences in their constant
regions)
• κ (kappa) : encoded on chromosome 2
• λ (lamda) : encoded on chromosome 22
– Both types can be found in all classes of immunoglobulins.
– Any one immunoglobulin molecule contains only one type.
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Structure of Immunoglobulin
• Heavy chains
– Distinct for each of the five classes of immunoglobulins.
• γ for IgG
• α for IgA
• μ for IgM
• ε for IgE
• δ for IgD
– All five types are encoded on chromosome 14.
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Structure of Immunoglobulin
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Structure of Immunoglobulin
Amino terminal end
VL
CL
Fab fragments
VH
CH1
CH2
Fc fragment
CH3
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IgG
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IgG
VL
CL
VH
CH1
CH2
CH3
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IgG
Amino terminal end
VL
CL
VH
CH1
CH2
Interchain disulfide
bonds
CH3
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IgG
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IgA
• It is the main immunoglobulin found in following secretions:
• Saliva
• Tears
• Respiratory tract secretions
• Intestinal tract secretions
• Genital tract secretions
• Colostrum
• Secretory IgA are dimers composed of two H2L2 units plus one unit of J
chain and secretory components.
• Serum IgA are monomers composed of single H2L2 unit.
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IgA
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IgA
J chain
Secretory component
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IgA
• Secretory Component:
– It is a polypeptide made by epithelial cells.
– It helps in passage of IgA to the mucosal surface and prevents its
degradation in the intestinal tract.
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IgM
• Main immunoglobulin for primary immune response.
• It is found as monomer on surface of all B lymphocytes where it acts
as antigen-binding receptor.
• It is found as pentamer in the serum composed of five H2L2 units
plus one J chain.
• IgM pentamer has 10 antigen binding sites.
• IgM can fix complement.
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IgE
• It can mediate immediate hypersensitivity reaction.
• It is the main defense against helminth (worm) infections (e.g.
Ascaris, Trichinella, Strongyloides) and hook worm infections
(Necators, Ancylostoma).
• Worms are too large for phagocytes to ingest. So they are killed by
worm-destroying enzymes released by eosinophils.
• IgE specific for worm protein binds to receptors on the surface of
eosinophils and trigger Antibody-dependent cellular cytotoxicity
(ADCC).
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IgD
• No known antibody function.
• Present on surface of many B lymphocytes.
• May act as antigen receptor.
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Important Functions of Antibodies
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Important Function of Antibodies
Antibody Function
IgG Opsonizes bacteria.
Fixes complement.
Neutralizes bacterial toxins and viruses.
Crosses the placenta.
IgA Secretory IgA prevents attachment of bacteria and viruses to
mucous membrane.
IgM Fixes complement.
Acts as antigen receptor on surface of B cells.
IgE Mediates immediate hypersensitivity.
Defends against worm infections.
IgD Uncertain.
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Polyclonal & Monoclonal Antibodies
• Polyclonal Antibodies:
– Formed by several different clones of plasma cells in response
to typical antigens.
– They are heterogeneous.
• Monoclonal Antibodies:
– Formed by single clone of plasma cells.
– They are homogeneous.
– Example: In plasma cell tumor (myeloma).
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Concept of Isotypes, Allotypes & Idiotypes
• Isotypes:
– Antigenic differences in their constant regions.
– All isotypes are found in all normal humans.
• Allotypes:
– Additional antigenic features of immunoglobulins that differ
among individuals.
• Idiotypes:
– Antigenic determinant formed by the specific amino acids in the
hypervariable region.
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Thank You !
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