Premium
A three‐phase pattern in growth, monoclonal antibody production, and metabolite exchange in a hybridoma bioreactor culture
Author(s) -
Bushell M. E.,
Bell S. L.,
Scott M. F.,
Snell K.,
Spier R. E.,
Wardell J. N.,
Sanders P. G.
Publication year - 1993
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260420118
Subject(s) - glutamine , metabolite , glutamine synthetase , ammonium , ammonia , biology , bioreactor , biochemistry , alanine , chemistry , amino acid , botany , organic chemistry
The use of partial cubic spline data interpolation for the calculation of volumetric metabolite exchange rates suggested the existence of three distinct metabolic phases during bioreactor culture of a hybridoma cell line. During phase 1, a rapid amino acid uptake rate and ammonia release rate were observed. The growth rate was low and glutamine synthetase activity fell. In phase 2, maximum growth rate and minimum glutamine assimilation and ammonium production rates were observed. Attempts to corroborate the apparent ammonia assimilation in this phase using 15 NH 4 Cl resulted in low incorporation rates into alanine and glutamine. Maximum glutamine synthetase activity took place during this period. Maximum antibody production rate was observed during phase 3 during which peaks in glutamine assimilation, ammonia release, and glutamine synthetase activity were observed. The apparent existence of the three phases prompted us to carry out Northern blot analysis of glutamine synthetase RNA at appropriate times during the process. This revealed a pattern of appearance and dis‐appearance of mRNA consistent with the three phases indicated by the fermentation parameters. © 1993 John Wiley & Sons, Inc.