Multiparametric analyses of hybridoma growth on glass cylinders in a packed-bed bioreactor system with internal aeration. Serum-supplemented and serum-free media comparison for MAb production.
Monoclonal antibodies are one of the most important products of biotechnology and laboratories and companies all over the world are pursuing their large-scale production. Herein we report a protocol for hybridoma cell cultivation over small glass cylinders inside a 3 liter bioreactor vessel which leads to the production and purification--in order of grams--of one MAb intended for human therapeutic use. This protocol proved to be simple, reproducible and cost effective. Three trials are reported: the first two using conventionally serum-supplemented medium culture and producing 3.15 and 2.1 g of purified MAb in 30 and 21 days respectively, and the third one using serum-free medium culture and producing 6 g of purified MAb in 36 days. We have ascertained the stability of the hybridoma by its cloning directly in serum-free medium. The downstream processing of the serum-free trial was done in a single step, concentrating large volumes of supernatant while simultaneously purifying the antibody.
Moro
A M
AM
Centro de Biotecnologia, Instituto Butantan, Sao Paulo, Brazil.
Rodrigues
M T
MT
Gouvea
M N
MN
Silvestri
M L
ML
Kalil
J E
JE
Raw
I
I
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D001769:Blood; D017252:CD3 Complex / Q000276:immunology*; D002455:Cell Division; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D016895:Culture Media, Serum-Free; D046508:Culture Techniques / Q000379:methods; D015194:Diffusion Chambers, Culture / Q000295:instrumentation; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000166:cytology*; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D010100:Oxygen
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D016895:Culture Media, Serum-Free; D007074:Immunoglobulin G; D010100:Oxygen