Strategies to extend longevity of hybridomas in culture and promote yield of monoclonal antibodies

Two-Stage Chemostat Studies of Hybridoma Growth, Nutrient Utilisation, and Monoclonal Antibody Production

“…Similar results to these have been obtained by Luan et al (1987) who extended the stationary phase of cell growth for several days, and thereby increased the final monoclonal antibody concentration, by adding essential amino acids, glutamine and vitamins. However, in the control culture growth was limited by an essential amino acid other than glutamine.…”

“…The supplementation of hybridoma cultures with extra essential amino acids has been reported to increase antibody titre in batch culture (Rupp, 1985;Luan et aL, 1987;Duval et aL, 1989 and. However, the specific role of individual amino acids in antibody production is not yet known, neither is it known whether the positive effect of supplementing batch cultures with extra amino acids has a positive effect on antibody production directly or indirectly through its effect on cell viability and growth rate.…”

Protein‐free fed‐batch culture of non‐GS NS0 cell lines for production of recombinant antibodies

“…This publication presents fed-batch culture data generated using an optimized second generation fedbatch process (FBP-2). The FBP development strategy was to increase the integral of viable cell concentration (IVCC) by further fortifying the nutrients present in the basal and feed media (deZengotita et al, 2000;DiStefano et al, 1996;Luan et al, 1987) above previous FBP-1 levels, and to take advantage of the efficient NS0 cell metabolism and higher cell line productivities while minimizing culture osmolality increase during the exponential growth phase (Ryu and Lee, 1999). Five NS0 cell lines, expressing five different antibodies, cultured in FBP-2 consistently achieved a volumetric productivity exceeding 120 mg/L per day.…”

Feeding of nutrients delays apoptotic death in fed-batch cultures of recombinant NSO myeloma cells

“…We have constructed cell lines that secrete antibody at high rates, typically from 15 to 50 pg/c-d, by transfecting NS0 myeloma cell lines with vectors expressing the selectable marker, glutamine synthetase (Bebbington et al, 1992), and the immunoglobulin heavy and light chain genes (for review see Robinson, et al, 1995). By analyzing the nutritional uptake of these cell lines and adding concentrated solutions of glucose, amino acids, and proteins, we have developed fed-batch processes that increase the specific secretion rate two-fold and the integrated number of viable cells (Luan et aL, 1987) five-fold, resulting in a ten-fold increase in the final product concentration (Robinson et al, 1994. ) These feeding strategies have resulted in final Ab yields approaching and exceeding 1 g/L (for review see Bibila and Robinson, 1995).…”