Cross flow diafiltration of serum with basal medium suitable for growth of hybridomas, sterilization and protein reduction.
Fetal Calf Serum (FCS) was extensively extracted by cross flow diafiltration (Pellicon, Millipore) and sterilized using the basal growth medium (DMEM) for the extraction. Ultrafiltration membranes of 10(5) and 3 X 10(5) Dalton cut off were used respectively. The diafiltrates were used for hybridoma cultivation and the results of growth and mAb-production were compared with standard medium (DMEM + 5% FCS). Slightly reduced mAb-titers were achieved. These were, however, compensated by decreased concentration of contaminating protein and higher specific mAb/protein ratio as examined by SDS-PAGE and enzyme linked immuno electro transfer blot (EITB).
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001709:Biotechnology / Q000379:methods; D003470:Culture Media; D003956:Dialysis; D005374:Filtration; D006825:Hybridomas / Q000166:cytology*; D051379:Mice
D000911:Antibodies, Monoclonal; D003470:Culture Media