Production of monoclonal antibodies in serum free medium.

We have succeeded in growing several established hybridoma cell lines (1116NS-19; 116NS-33a; 1116NS-52a; 1083-17-1A; 691-19-19) in a serum free medium supplemented with physiological concentrations of insulin (5 microgram/ml) and transferrin (5 microgram/ml). The hybridoma cells replicate in this medium and will secrete monoclonal antibodies in quantities comparable to those produced in the presence of serum. Adapted hybridoma cultures have been secreting antibodies in the supplemented serum free (SSF) medium for more than 3 months. Parental mouse myeloma P3x63Ag8 cells and non-secreting subclone 653 cells cannot survive longer than 6 days under the same conditions. Both insulin and transferrin are necessary for hybridoma cells to grow in serum free medium and omission of either one will not permit cell multiplication.