Hybridoma Technology

                                   Hybridoma Technology 

Hybridoma Technology is the technology in which hybrid cells are produced by fusing a specific antibody producing B-cell with myeloma cell . The process start  by injecting a specific antigen  into a mouse and collect the antibodies producing cell from the spleen and fuse with the myeloma cells . Both of this fusing partner give hybrid cell known as Hybridoma cells . Now these cells are transferred to a medium in which the myeloma cells die , but the hybridomas survive . Therefore these cells are transferred  into HAT medium and aminopterin are also added . The work of aminopterin is to block de novo synthesis . Normally the cells are survive in this medium but if the cell are unable to  produce the HGPRT medium then they are unable to use de novo pathway and die in aminopterin containing medium .  After fusion of lymphocytes with HGPRT - negative myeloma  cells in an aminopterin containing medium which is supplemented with hypoxanthine and thymidine to ensure sufficient supply of substrate for the salvage pathway is added which kills myeloma cells but allow hybridomas to survive and in this way monoclonal antibodies are produced .