TAILIEUCHUNG - HYBRIDOMA TECHNOLOGY FOR PRODUCTION OF MONOCLONAL ANTIBODIES

Hybridomas are cells that have been engineered to produce a desired antibody in large amounts. To produce monoclonal antibodies, Bcells are removed from the spleen of an animal that has been challenged with the relevant antigen. These B-cells are then fused with myeloma tumor cells that can grow indefinitely in culture (myeloma is a B-cell cancer). This fusion is performed by making the cell membranes more permeable. The fused hybrid cells (called hybridomas), being cancer cells, will multiply rapidly and indefinitely and will produce large amounts of the desired antibodies. They have to be selected and subsequently cloned by limiting dilution. Supplemental media containing Interleukin-6. | Volume 1 Issue 2 March - April 2010 Article 017 ISSN 0976 - 044X HYBRIDOMA TECHNOLOGY FOR PRODUCTION OF MONOCLONAL ANTIBODIES Shivanand Pandey Smt. R. B. P. M. Pharmacy College Atkot-360040 Rajkot Gujarat. India Email ABSTRACT Hybridomas are cells that have been engineered to produce a desired antibody in large amounts. To produce monoclonal antibodies B-cells are removed from the spleen of an animal that has been challenged with the relevant antigen. These B-cells are then fused with myeloma tumor cells that can grow indefinitely in culture myeloma is a B-cell cancer . This fusion is performed by making the cell membranes more permeable. The fused hybrid cells called hybridomas being cancer cells will multiply rapidly and indefinitely and will produce large amounts of the desired antibodies. They have to be selected and subsequently cloned by limiting dilution. Supplemental media containing Interleukin-6 such as briclone are essential for this production of monoclonal anti-bodies was first invented by Cesar Milstein Georges J. F. Kohler and Niels Kaj Jerne in 1975. Selection occurs via culturing the newly fused primary hybridoma cells in selective-media specifically media containing 1x concentration HAT for roughly 10-14 days. After using HAT it is often desirable to use HT containing media. Cloning occurs after identification of positive primary hybridoma cells. Clone by limited dilution. While some may believe that IL-6 is essential for this step it is not necessary to add that expensive supplement rather use 50 heat-inactivated FBS for the first week. Add 10 FBS DMEM to the clone culture plate after screening for single colony wells. Keywords Hybridomas monoclonal antibodies Interleukin-6 Supplemental media INTRODUCTION Hybridomas are cells that have been engineered to produce a desired antibody in large amounts to produce monoclonal antibodies. 1 2 Monoclonal antibodies can be produced in specialized cells through a technique .

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