Characteristics of HAb18 Cell Growth， Cell Metabolism and Antibody Synthesis under Serum Free Culture Condition

Abstract: A serum free medium for hybridoma HAb18 was obtained through decreasing the serum concentration gradually， analyzing amino acids exhaustion with HPLC and screening serum supplement through orthogonal experiment. Cell grown in serum free medium was subjected to cell counting. Supernatant from the spent medium were pooled to determine the concentration of glucose， glutamine， lactate and ammonia. Moreover， not only antibody yield and antigen binding activity were detected， but dynamic parameters were analyzed. Our results showed that the maximum cell density and antibody production in serum free medium were 0.91?06 cells/ml and 43.8 mg/L， respectively. Although the specific cell growth rate reduced slightly， antibody productivity increased (0.0207/h vs. 0.0218/h， 0.387 vs. 0.218 pg/cell/h， P<0.01). Glucose and glutamine exhaustions in serum free medium were similar to that in serum containing medium; while lactate production cut down and ammonia production enhanced. Likewise， antigen-binding activity increased. Taken together， studies on characteristics of HAb18 cell growth， metabolism and antibody synthesis in serum free medium would lay a foundation for HAb18 serum free suspension fed-batch technology.

CSTR: 32200.14.cjcb.2006.01.0018