Three-dimensional culture for monoclonal antibody production by hybridoma cells immobilized in macroporous gel particles.
Cell proliferation and long-term production of monoclonal antibody IgG(2b) by M2139 hybridoma cells immobilized in macroporous gel particles (MGPs) in packed-bed reactor were studied for a period of 60 days. The MGPs were made of supermacroporous gels produced in frozen conditions from crosslinked polyacrylamide and modified with gelatin which were housed in special plastic carriers (7 x 9 mm(2)). Cells were trapped in the interior part of MGPs by attaching to the void space of the gel matrix as three-dimensional (3D) cultivation using gelatin as a substrate layer. Optimizing productivity by hybridoma cell relies on understanding regulation of antibody production. In this study, the behavior of M2139 cells in two-dimensional cultures on multiwell plate surfaces was also investigated. The effect of three different medium such as basal medium Dulbecco's modified Eagle's medium (D-MEM) containing L-glutamine or L-glutamine + 2 mM alpha-ketoglutarate or L-alanyl-glutamine (GlutaMAXtrade mark) was studied prior to its use in 3D cultivation. The kinetics of cell growth in basal medium containing L-glutamine + alpha-ketoglutarate was similar to cells grown on GlutaMAX containing medium, whereas D-MEM containing L-glutamine showed lower productivity. With the maximal viable cell density (6.85 x 10(6) cells mL(-1)) and highest specific mAb production rate (3.9 mug mL(-1) 10(-4) viable cell day(-1)), D-MEM-GlutaMAX was further selected for 3D cultivation. Cells in MGPs were able to grow and secrete antibody for 30 days in packed-bed batch reactor, before a fresh medium reservoir was replaced. After being supplied with fresh medium, cells again showed continuous growth for another 30 days with mAb production efficiency of 50%. These results demonstrate that MGPs can be used efficiently as supporting carrier for long-term monoclonal antibody production.
Nilsang
Suthasinee
S
Dept of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, Lund, Sweden.
Nehru
Vishal
V
Plieva
Fatima M
FM
Nandakumar
Kutty Selva
KS
Rakshit
Sudip Kumar
SK
Holmdahl
Rikard
R
Mattiasson
Bo
B
Kumar
Ashok
A
D000180:Acrylic Resins / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002460:Cell Line; D049109:Cell Proliferation; D018914:Cells, Immobilized / Q000166:cytology / Q000378:metabolism; D005780:Gelatin / Q000737:chemistry; D005782:Gels / Q000737:chemistry; D006825:Hybridomas / Q000166:cytology / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D010316:Particle Size; D016062:Porosity; D013499:Surface Properties; D013997:Time Factors; D000099166:Polyacrylamides
D000180:Acrylic Resins; D000911:Antibodies, Monoclonal; D005780:Gelatin; D005782:Gels; D000099166:Polyacrylamides