B-cell cultivation method

A method for co-cultivating a single peripheral blood B-cell for production of an antibody, wherein said B cell is deposited into a well of a multi-well plate using fluorescence activated cell sorting (FACS) comprising the steps of:

• a) supplementing a co-cultivation medium with a feeder mix, the feeder mix consisting of additives promoting the activation or survival of B cells or antibody secretion,
• wherein the co-cultivation medium supplemented with the feeder mix contains no other additives promoting the activation or survival of B cells or antibody secretion but those of feeder mix;
• and
• b) co-cultivating the B-cell with feeder cells in the presence of the co-cultivation medium supplemented with the feeder mix,
• wherein neither of a thymocyte cultivation supernatant, a macrophage cultivation supernatant, or a T-cell cultivation supernatant is added to the co-cultivation medium supplemented with the feeder mix,
• wherein the final concentration of the additives promoting the activation or survival of B cells or antibody secretion in the co-cultivation medium supplemented with the feeder mix is:
• 0.05-0.35 ng/ml phorbol myristate acetate (PMA),
• 0.02-0.2 ng/ml IL-1beta,
• 0.02-0.2 ng/ml TNFalpha,
• 0.5-5 ng/ml IL-2,
• 0.1-1 ng/ml IL-10,
• 0.1-1 ng/ml IL-6;
• and
• wherein said B-cell produces an antibody.