Why Is Hybridoma Technology So Important?

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The first description of the hybridoma technology was appeared in 1975 and then it seemed to hold enormous promise for the treatment of a variety of human disease. The ability to produce monoclonal antibodies led to the availability of large amounts of homogeneous and predictable preparations of antibody. The potential to renew indefinitely a particular antibody surmounted many of the technical and regulatory problems that made polyclonal antisera difficult to use as therapeutic agents in man.

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The hybridoma technology seemed even more valuable as it became clear that it could lead to the generation of pure, highly specific antibodies from impure, poorly characterized antigens. Monoclonal antibodies have been extremely useful in basic investigations and have facilitated the development of new diagnostic tests for serum and tissue components and infectious agents.

Application The hybridoma technology is used to produce

monoclonal antibodies.http://www.creativebiomart.net/recombinantproteins-Hybridoma.htm

Procedure of Hybridoma Technology

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B-lymphocytes are extracted from the spleen of an animal, but usually it is extracted from the mouse, which has been immunized with the required antigen which monoclonal antibodies are produced. Mouse is immunized by giving antigen injection along with an adjuvant via subcutaneously or by peritoneal cavity; this is followed by booster doses of the antigen. Adjuvant is nonantigenic in nature but they stimulate the immune system.

This immunization with specific antigen increases the specific antibody producing B-lymphocytes; this considerably increases the chances of obtaining the required hybridoma cells or clones.

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This specific antibody producing B-lymphocytes are then mixed with the seleceted myelma cells and are induced to fuse to form hybrid cells. They myeloma cells are selected based on some criteria like these cells themselves should not produce antibodies and also they should contain a genetic markers such as HGPRT. This genetic marker helps in easy selection of the resulting hybrid cells.

When HGPRT myeloma cells are fused with specific antibody producing B-lymphocytes, the resulting cell population will have the mixture of cell population such as hybrid cells, myeloma cells, B-lymphocytes.

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This mixture of cell population is then cultured in selective media known as HAT medium along with the drug aminopterin. The specific antibody producing B-lymphocytes are unable to divide continuously in the culture medium, therefore eventually they die.

Only the hybridoma cells have got the ability to divide and proliferate on the HAT medium because genome from the B-lymphocyte makes them HGPRT positive and genome from the myeloma cells they can divide indefinitely. Thus only the hybridoma cells or fused cells are selected using selective media called as HAT medium.

http://www.creativebiomart.net/recombinantproteins-Hybridoma.htm

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