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Molecular Biology - Essay Example

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Monoclonal antibodies are used in research, diagnostics and therapeutics. Monoclonal antibodies are issued from one single clone of cells. Monoclonal antibodies from mouse origin are easy to produce but their therapeutic availability is restricted by their antigenicity…
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Molecular Biology
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"Molecular Biology"

Download file to see previous pages For monoclonal antibody, mice are immunised with purified protein.
Mouse splenocytes are collected after the immunisation process and fused with myeloma cells. To obtain monoclonal antibodies individual B cells are fused to myeloma cells and isolated by serial dilution resulting to a fusion product or hybridoma cell line of which can produce one specific antibody for extended periods of time by tissue culture. These hybridomas can be screened and best clones cultured in standard tissue culture facilities. The hybridoma cell line obtained for a given antibody was injected into the peritoneal cavity of mice where it grew and simultaneously produces the antibodies. Ascetic fluid containing the antibodies was harvested from the peritoneal cavity after a period of time.
One fusion may produce 1000hybridomas therefore one must choose the most appropriate. This might be the highest infinity mAb but could be the most stable, the least cross reacting, the highest specificity depending on what is required.
The advantage of using monoclonal antibodies is that they are monospecific, they tend to reduce cross reactivity and useful in diagnostics such as tissue typing. They are highly reproducible, can use relatively impure antigens to immunise animas, theoretically have limitless supply and can manufacture using recombinant DNA technology plus phage display libraries to produce fully human antibodies of any specificity.
Labelling antibodies is useful as it enables detection of antibodies. Currently, the most commonly used are fluorescent labels, enzyme labels, chemiluminescent labels and radioactive labels. These are sometimes amplified using Avidin-Biotin Conjugate system. Some common fluorescent labels include, Fluorescein isothiocyanate, DAPI, Phycoerithrin and Texas red.
When using enzyme labels, an enzyme is covalently attached to the ...Download file to see next pagesRead More
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