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Antibodies & Antigens Pin Ling (凌 斌), Ph.D.
ext 5632; References: 1. Abbas, A, K. et.al, Cellular and Molecular Immunology (6th ed., 2007), Chapter 4 2. Male D., J. Brostoff, D. B Roth, and I. Roitt Immunology (7th ed., 2006), Chapter 3
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Question What effect you expect on the immune system if the thymus was
removed? Ans: 1. Total lymphocytes are drastically reduced. T cell development was blocked. B cells are also reduced => require T helper cells for their proliferation. LN size is reduced. => Get infections easier. 2. DiGeorge Syndrome => patients w/ congenital thymic aplasia => Fewer T cells in defected thymus
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Outline The origin concept of Antibodies
Structures & Features of Antibodies Antibody binding of Antigens Applications of Antibodies Summary & Question
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Definition of Key Terms
1. Antigen (Ag) - A molecule can specifically binds to an Antibody or a T-cell receptor (TCR) and usually induces an adaptive immune response. - Antigen Determinant (Epitope): The specific portion of an Ag recognized by an Ab or TCR. 2. Antibody (Ab) - Also called immunoglobulin (Ig), A type of glycoprotein produced by B cells that binds Ags with high specificity & affinity. - Binds to Ags including all classes of molecules, ex. Protein, Lipid, Carbohydrate, or Chemical. 3. Antiserum - Serum from an Ag-immunized individual that contains Ab specific Ag
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In 1890, Behring & Kitasato => Serum therapy
Inactivated toxin => Animal-A => Protective immunity Serum from Animal-A => Animal-B => Passive immunity Serum A contains “Anti-toxin” proteins => Humoral immunity “Anti-toxin” => “Anti-body” that recognize many other substances in addition toxin.
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Outline Structures & Features of Antibodies
The origin concept of Antibodies Structures & Features of Antibodies Antibody binding of Antigens Applications of Antibodies Summary & Question
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Structures of Antibodies-I
Antibody (Immunoglobulin, Ig) 1. Two identical Light chains Two “ “ Heavy chains 2. Each chain has repeating unit, Ig domain 3. Chains are linked by disulfide bonds 4. Each chain consists of - Variable region (N-terminus) - Constant region (C-terminus)
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Structures of Antibodies-II
Ag-binding site Antibody (Immunoglobulin, Ig) 5. Share basic structure features 6. Show the remarkable variability in regions for Ag bindings => H-Variable region + L-Variable region =>Antigen binding site 7. Heavy chain Constant region => Effector functions Effector functions
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Proteolytic cleavage of Antibody
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Ab Isotypes-I 1. 5 distinct classes: Ig A, D, E, G, & M
Some => subclasses Heavy chain => Classification 2. Different Ig isotypes => Different Effector functions 3. IgG => predominant & long half-life IgA => Mucosal lumens & milk 4. Membrane-bound Ig M & D => B-Cell antigen Receptor (BCR)
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Ab Isotypes-II
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Ab Isotypes-III
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Key Concepts in Ab isotypes
1. IgM is the predominant Ab during the primary immune response and also functions as a BCR. 2. IgG is the predominant Ab during the secondary immune response. 3. IgA is produced in 2nd immune response and plays a key role in mucosa immunity area (ex. Respiratory & GI tracts). 4. IgD is a membrane-bound Ag receptor on B cells. 5. IgE have evolved to protect against helminth parasites. 6. Fc receptors expressed on various immune cells => Mediate Ab effector functions
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Structures of Ab Isotypes
They differ in: Size Charge Amino acid seq Carbohydrate content
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Membrane and secreted forms of Abs
IgD => only membrane form
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Ig expression during B cell maturation & activation
Membrane-bound IgD or IgM => BCR + Ag => B cell activation => Plasma cells => Secreted Abs
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Changes in Ab structure during humoral immune responses
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Ab-mediated Immune Effector Systems
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Secreted IgA dimer => Mucosal lumen
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Outline Antibody binding of Antigens The origin concept of Antibodies
Structures & Features of Antibodies Antibody binding of Antigens Applications of Antibodies Summary & Question
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Key Concepts in Ab-Ag interaction
1. Antibody (Ab) form multiple non-covalent bonds with antigen=> Reversible - Attractive forces (H bonds, electrostatic bonds, van der Waals forces, & hydrophobic forces) => High affinity interaction 2. The Ag-binding sites of an Ab are complementary to the conformation of Ag determinants (epitopes) of an Ag. 3. Affinity vs. Avidity for AbAg Affinity => A measure of the strength of interaction between an Ag-binding site and its epitope => Kd, dissociation constant; small Kd => stronger affinity Avidity => The overall strength of AbAg => Affinity & the valency of interactions
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Hypervariable Regions form the Ag-binding site
Most of sequence differences among Abs => Three short stretches in V regions => Hypervariable regions (HV), also called Complementarity- determining regions (CDRs)
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Complementary interactions between Ag-binding sites and their epitopes
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Specificity, Cross-reactivity & non-reactivity of AbAg
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The Nature of Ag determinants
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Antibody & other Antigen-Recognizing Molecules
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Valency and Avidity of AbAg interactions
A pentameric IgM => low-affinity for each valent binding => Many low-affinity binding => high avidity interaction
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Flexibility of AbAg interactions
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Outline Applications of Antibodies The origin concept of Antibodies
Structures & Features of Antibodies Antibody binding of Antigens Applications of Antibodies Summary & Question
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The Development of monoclonal Ab
Unlimited production of unique Ab for a specific Ag => Revolutionize Immunology & other fields Its Applications: Identification of phenotypic markers Immunodiagnosis & Immunotherpy Tumor diagnosis & therapy
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The Development of monoclonal Ab-II
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Immunodiagnosis-ELISA
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Immunoprecipitation
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Outline Summary & Question The origin concept of Antibodies
Structures & Features of Antibodies Antibody binding of Antigens Applications of Antibodies Summary & Question
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SUMMARY 1. All Abs have s common symmetric structure:
(2 Heavy chains + 2 Light chains ) meanwhile show the remarkable variability in regions for Ag bindings 2. Abs are classified into different isotypes on the basis of different Heavy chain C regions. Ab-mediated effector functions also depend on the H-chain C regions. 3. Five classes of Ab in mammals: IgA, IgD, IgE, IgG & IgM 4. Monoclonal Abs are applied to many fields for research & clinical treatment.
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Question What mechanisms to achieve the generation of Ab diversity?
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