Preparation of Antibodies

by Peter Delves, PhD

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    Antibodies are incredibly useful in a number of different diagnostic assays and actually throughout the whole of biology, there are uses for monoclonal antibodies because their really high degree of specificity makes them excellent probes to detect the presence of particular molecules. One can produce polyclonal antibodies, that is antibodies produced from a number of different B-cells by immunizing an animal with an antigen. So here we have an example, taking a protein and immunizing a sheep with this particular protein. One can then collect blood from the sheep a period of time later, one gives the sheep a chance to make some antibodies, the immune response gets going. And then a few weeks later, after a couple of boosters, one can take some blood from the sheep. And there will be polyclonal antibodies that are specific for this antigen. And the proportion of antibodies because you’ve immunized the sheep, the proportion of antibodies against this particular antigen will be increased. So they won’t all be against this antigen, there’ll be antibodies against many other antigens as well. But the percentage of antibodies against the immunizing antigen will have been increased by the immunization procedure. So you’ll have different antibodies against different parts of the particular antigen of interest. So for example, immunizing with human IgG, you can develop sheep anti-human IgG as a reagent to use in immunodiagnostic tests, where you want to see if a patient has IgG antibodies that bind to a particular antigen. For example, an autoantigen. And using these kind of sheep antibodies and then labeling them with a fluorescent dye or with an enzyme will allow their use in immunodiagnostic assays. So those are polyclonal antibodies, a mixture of antibodies. And very often it’s useful to have a mixture of antibodies rather than a...

    About the Lecture

    The lecture Preparation of Antibodies by Peter Delves, PhD is from the course Immunodiagnostics. It contains the following chapters:

    • Production of Polyclonal Antibodies
    • Hybridoma Production of Monoclonal Antibody
    • Phage Display Production of Monoclonal Antibody

    Included Quiz Questions

    1. Myeloma cell line
    2. Melanoma cell line
    3. Bacteriophages
    4. B-cell stimulating cytokines
    5. Fusion of normal T-cells
    1. B cell antigen specificity and myeloma cell immortality
    2. B cell antigen specificity and myeloma cell splenic specificity
    3. B cell splenic specificity and myeloma cell immortality
    4. B cell immortality and myeloma cell splenic specificity
    5. B cell immortality and myeloma cell antigen specificity
    1. IGHV and IGLV
    2. mRNA
    3. Flexible linker
    4. Tumor antigen
    5. Fused myeloma and B cells
    1. Antibody on cell surface and gene for encoding that antibody within cell
    2. Identical antibodies on cell surfaces
    3. Two IGHV segments linked with a flexible linker
    4. mRNA expression for various antibodies
    5. Specific primers that amplify cDNA

    Author of lecture Preparation of Antibodies

     Peter Delves, PhD

    Peter Delves, PhD

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