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

J Immunol Methods. 1994 Nov 10;176(1):67-77. doi: 10.1016/0022-1759(94)90351-4.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / isolation & purification
  • Blood*
  • CD3 Complex / immunology*
  • Cell Division
  • Cells, Cultured
  • Chromatography, Affinity
  • Culture Media, Serum-Free*
  • Culture Techniques / methods
  • Diffusion Chambers, Culture / instrumentation
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Hybridomas / cytology*
  • Immunoglobulin G / immunology
  • Mice
  • Oxygen

Substances

  • Antibodies, Monoclonal
  • CD3 Complex
  • Culture Media, Serum-Free
  • Immunoglobulin G
  • Oxygen