Production of monoclonal antibodies after in vitro immunization

Production of monoclonal antibodies after in vitro immunization 

Spleen cells are obtained from an unprimed BALB/c mouse (Section 

washed and diluted to 20 ml with medium containing about 1 pg of immunogen per ml or 10 irradiated (2500 rad) or ethanol-fixed cells. Then, 10 ml of thymocyte-conditioned medium (Section 

5) is quickly thawed and added to the spleen cell suspension which is mixed and left undisturbed for 5 days in a 75 cm tissue culture flask in a 5% CO2 atmosphere. After 5 days, when numerous blast cells are visible, the cells are ready for fusion with myeloma cells (line 653 is recommended). Fusion is performed as described in Section 5.4.3.1. 

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