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7 Cards in this Set
- Front
- Back
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What are the 2 basic functions of antibodies |
1. N terminal region binds to epitopes of anigen 2. C terminal region or constant region recruits cell to kill pathogen and is reponsbible for the effector functions |
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how are light and heavy domains made |
light chain has V and L regions which are splice and rearranged heavy chain V D J regions which are spliced and rearranged more varibaliity in the heavy chain domain light chain join randomly with heavy chains 10 ^11 possible rearrangement for the variable region |
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How are antibodies assembled |
affinity maturation - B cells will produce antibodies with increase affinitiy to be kept alive B T cells in the process of clonal selection HV hotspots are prime for mutation which can lead to better or worse affinity to epitope of antigen |
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What were early approaches to antibodies |
immunize with antigen collect antiserum containing only proteins and ab no RBC or clotting proteins ab will then be isolated via MW/bead affinity chromatorgraphy once antisera is used ab cannot be regenerated |
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hybridoma |
1. boost mouse with antigen and adjuvants to boost the immune system resulting in a greatest production of lymphocytes 2. remove spleen of mouse and collect all B cells 3. combine B cells with HGPRT-ve myseloma cells in a PEG solution to melt/fuse membrane 4. Place through HAT selection 5. conduct elisa on hybridoma and dilute to create 0.3 cells per uL in a well thus once cell per well. 6. Add feeder cells in the 96 well plate and watch monoclonal antibodies proliferate |
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What is papain used for |
cuts above disulfide bond
avidity is reduced by 1/2 FC region is still function but binds with C1q |
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pepsin |
cuts below disulfide bond avidity are the same FC is however not functional |
