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    MBBS Hybridoma Technology

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    Martins Ahiakwo
    This document discusses hybridoma technology and monoclonal antibodies. It explains that monoclonal antibodies are identical antibodies produced by a single parent cell, allowing them to target a specific antigen. The technique of producing monoclonal antibodies was discovered in 1975 and involves fusing antibody-producing B cells with myeloma cells to generate stable hybridoma cell lines that produce monoclonal antibodies of a single specificity. This hybridoma technology allows for the mass production of monoclonal antibodies that have various applications in medicine and research.

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    MBBS Hybridoma Technology

    Uploaded by

    Martins Ahiakwo
    109 views10 pages
    This document discusses hybridoma technology and monoclonal antibodies. It explains that monoclonal antibodies are identical antibodies produced by a single parent cell, allowing them to target a specific antigen. The technique of producing monoclonal antibodies was discovered in 1975 and involves fusing antibody-producing B cells with myeloma cells to generate stable hybridoma cell lines that produce monoclonal antibodies of a single specificity. This hybridoma technology allows for the mass production of monoclonal antibodies that have various applications in medicine and research.

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    MBBS Hybridoma Technology

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    This document discusses hybridoma technology and monoclonal antibodies. It explains that monoclonal antibodies are identical antibodies produced by a single parent cell, allowing them to target a specific antigen. The technique of producing monoclonal antibodies was discovered in 1975 and involves fusing antibody-producing B cells with myeloma cells to generate stable hybridoma cell lines that produce monoclonal antibodies of a single specificity. This hybridoma technology allows for the mass production of monoclonal antibodies that have various applications in medicine and research.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    109 views10 pages

    MBBS Hybridoma Technology

    Uploaded by

    Martins Ahiakwo
    This document discusses hybridoma technology and monoclonal antibodies. It explains that monoclonal antibodies are identical antibodies produced by a single parent cell, allowing them to target a specific antigen. The technique of producing monoclonal antibodies was discovered in 1975 and involves fusing antibody-producing B cells with myeloma cells to generate stable hybridoma cell lines that produce monoclonal antibodies of a single specificity. This hybridoma technology allows for the mass production of monoclonal antibodies that have various applications in medicine and research.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 10

    Hybridoma Technology and

    Monoclonal Antibodies In
    Medicine and Biological
    Research.
    ANTIBODY
    An antibody is a protein used by the immune system to identify and
    neutralize foreign objects like bacteria and viruses. Each antibody
    recognizes a specific antigen unique to its target.

    Monoclonal antibodies (mAb) are antibodies that are identical


    because they were produced by one type of immune cell, all clones of
    a single parent cell.

    Polyclonal antibodies are antibodies that are derived from different


    cell lines.
    Monoclonal Antibodies
    Monoclonal antibodies (mAb) are antibodies that are
    identical because they were produced by one type of
    immune cell, all clones of a single parent cell. Given
    (almost) any substance, it is possible to create
    monoclonal antibodies that specifically bind to that
    substance; they can then serve to detect or purify that
    substance. This has become an important tool in
    biochemistry, molecular biology and medicine.
    The structure of antibody
    HYBRIDOMA TECHNOLOGY Georges Kohler

    It is technique of producing hybrid cell lines called


    hybridomas by the fusion of fusing a specific
    antibody-producing lymphocyte B cell with a Cesal Milstein
    myeloma cell that has an ability to grow in tissue culture
    Hydridoma produce antibodies produced that have
    single specificity and are called monoclonal antibodies.
    This technique was discovered Georges Kohler of West
    Germany, Cesal Milstein of Argentina and Niels Jerne of Niels Jerne
    Denmark in 1975.
    They were awarded Nobel Prize for Physiology and
    Medicine the 1984
    Method

    1. Immunization of a mouse
    2. Isolation of B cells from the spleen
    3. Cultivation of myeloma cells
    4. Fusion of myeloma and B cells
    5. Separation of cell lines
    6. Screening of suitable cell lines
    7. in vitro (a) or in vivo (b) multiplication
    8. Harvesting
    Steps in monoclonal antibody production by hybridoma
    technology

    1.Immunize a rabbit through repeated injection of a specific antigen for the


    production of specific antibody, facilitated due to proliferation of the desired B
    cells.
    2.Produce tumors in a mouse or a rabbit.
    3.Culture separately the spleen cells that produce specific antibodies and the
    myeloma cells that produce tumors
    4.Myeloma cells cannot synthesize antibodies as they lack HGPRT gene
    required for the synthesize the enzyme, hypoxanthine guanine phosphoribosyl
    transferase
    Fusion
    Fusion of spleen cells to myeloma cells is induced
    using polyethylene glycol (PEG), to produce
    hybridoma
    Hybridomas are grown in selective hypoxanthine
    aminopterin thymidine (HAT) medium.
    HAT medium contains a drug, aminopterin that
    blocks one pathway for nucleotide synthesis,
    making the cells dependent on another pathway
    that needs HGPRT enzyme, which is absent in
    myeloma cells.
    Myeloma cells that do not fuse with B cells will
    die.
    B cells that do not fuse will also die because they
    lack tumorigenic property of immortal growth.
    Selection and Storage
    Select desired hybridoma for cloning and
    antibody production
    Prepare single cell colonies that can grow and
    use them to screen of antibody producing
    hybridomas
    Only one in several hundred cell hybrids will
    produce antibodies of the desired specificity
    Culture selected hybridoma cells for the
    production of monoclonal antibodies in large
    quantities
    Hybridoma cells can be frozen for future use or
    can be injected in the body of an animal, so that
    antibodies will be produced in the body and
    recovered later from the body fluid.
    Applications

    DIAGNOSIS

    THERAPY

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