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23 Cards in this Set
- Front
- Back
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What cell produces antibodies? |
B-cells |
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What is the function of plasma cells in the immune system? |
Once activated, they can produce their own antibodies of their own |
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How does a CD4+ cell induce an antibody response? |
1) CD4+ (T-cell) surface receptor and CD28 binds B-cell MHC-Peptide complex and B7 respectively 2) B-cell binding stimulates T-cells to produce Cytokine and CD40 ligands which are bound by the B-cell 3) Binding stimulates differentiation of B-cell and proliferation of an appropriate antibody |
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How does a B-cell produce a MHC Class II-peptide complex? |
1) Binding of Microbial-bound Antigen to B-cell-bound receptor 2) Receptor mediated endocytosis of invader and upregulation of B7 receptor production 3) Digestion of invader into smaller peptides 4) Presentation of invader's antigen on MHC II = MHC-peptide complex on B-cell surface |
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What is the purpose of CD28? |
CD28-B7 binding stimulates production of cytokines |
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What is the product of B-cell proliferation? |
Memory and plasma cell formation |
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How do B-cell antibodies differ from plasma cell antibodies? |
Plasma antibodies are alternatively spliced compared to B-cells to exclude a transmembrane binding region |
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Define Fab and Fc domains |
Fab = Antigen Binding Fragment Fc = Crystallizable Fragment
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Describe the structure of an Antibody |
2 Fab and 1 Fc domain 2 identical Heavy chains that extend the entire length of the antibody 2 identical Light chains that only span the Fab domain All chains are linked by disulfide linkages |
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Describe the structure of the Fab domain |
Fab is linked to the Fc by a hinge region From the hinge region, there is a Constant H and L domain attached to a variable H and L domain attached to a Hypervariable H and L domain N-terminus at hypervariable tips |
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Describe Antigen/antibody binding |
Strong non-covalent interactions with dissociation constants in the 10^-7 to 10^-10 M range |
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How do Polyclonal and Monoclonal antibodies differ? |
Polyclonal have multiple epitopes, are produced by multiple B cell, and produce multiple antibodies Monoclonal only bind 1 epitope and are produced from 1 antibody |
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Difference between cellular and humoral response |
Humoral: Helper T-Cells (CD4+) scan for MHC Class II-peptide complexes and help stimulate production of antibodies Cellular: Killer T-cells scan for MHC Class I-peptide complexes and help stimulate proliferation of other phagocytizing Killer T-cells |
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How can we enrich the amount of antibody that recognizes our antigen? |
Affinity purification |
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How do we make a monoclonal antibody? |
Innoculate a mouse, wait, culture its spleen. Test if spleen cells have an antibody against your target antigen, and culture those that express the antibody. Repeat! |
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How can antibodies act as powerful detection reagents? |
1) To detect presence of antigen (western blots) 2) To measure antigen concentration 3) To purify antigen 4) To localize antigens in cells and tissue |
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How do we visualize antigen/antibody complexes? |
Conjugate the antibody to horseradish peroxidase (HRP) enzyme Upon reaction of antibody/antigen binding, the HRP has quantifiable chemiluminescence |
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What are the 2 main uses of ELISA? |
1) Can test multiple sample for presence of specific protein 2) Quantitative assays |
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How does the well differ in Sandwich and Indirect ELISA? |
Indirect: Antibody binds to antigen-coated well Sandwhich: Antigen binds to antibody-coated well |
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How does the enzyme-linked antibody binding differ between indirect and sandwich ELISA? |
Indirect: Enzyme-linked antibody binds to Fc region of antigen-bound antibody Sandwich: Enzyme linked polyclonal antibody binds to the antigen that was bound to the immobilized monoclonal antibody |
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What ELISA method do we use to detect HIV and PSA respectively? |
HIV: Indirect PSA: Sandwich |
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How do antibodies aid in membrane protein crystallization |
Binding antibodies to a membrane protein increases the amount of soluble domain thus increasing crystal contact areas and locking proteins in a specific state |
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What is herceptin |
A monoclonal antibody targeted to a cancer-cells EGF receptor (HER2 receptor) to prevent proliferation |
