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Hybridoma Technology - Coursework Example

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The paper "Hybridoma Technology" states that tissues and tumours can be classified based on their expression of certain markers, with the help of monoclonal antibodies.  They help in distinguishing morphologically similar lesions and in determining the organ or tissue origin of metastases…
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Hybridoma Technology
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Download file to see previous pages The advantage of the process is that it can combine the qualities of the two different types of cells; the ability to grow continually and to produce large amounts of pure antibody.
The use of monoclonal antibodies is numerous and includes the prevention, diagnosis, and treatment of disease. For example, monoclonal antibodies can distinguish subsets of B cells and T cells, which is helpful in identifying different types of leukaemias and lymphomas. Monoclonal antibodies are being used to track cancer antigens, to attack cancer metastases, and preventing graft-versus-host disease in bone marrow transplants.
The other applications of monoclonal antibodies include the use in immuno-histochemistry for the localization of specific antigens in situ, immunogold labelling, Western-type blotting, radioimmune assays, ELISA, and functional assays.
After the injection of a specific antigen into a mouse and procuring the B cells from the mouse's spleen, the splenic B cells are mixed with HGPRT (hypoxanthine-guanine phosphoribosyltransferase) negative myeloma cells and polyethene glycol (a fusing agent) before being fused with histocompatible myeloma cells like Sp2/0.
The mixing and centrifugation process generates myeloma–splenic B cell hybridomas, which are plated into tissue culture wells. After this, the unfused myeloma cells are removed by using a selective medium containing hypoxanthine, aminopterin, and thymidine (HAT). Removal of the unfused myeloma cells is necessary because they have the potential to outgrow other cells, especially weakly established hybridomas. Once a hybridoma colony is established, it will continually grow in culture medium like RPMI-1640 (with antibiotics and foetal bovine serum) and produce antibody.
The next stage is a rapid primary screening process, which identifies and selects only those hybridomas that produce antibodies of appropriate specificity. ...Download file to see next pagesRead More
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