Metabolic rates, growth phase, and mRNA levels influence cell-specific antibody production levels from in vitro-cultured mammalian cells at sub-physiological temperatures

Mol Biotechnol. 2008 May;39(1):69-77. doi: 10.1007/s12033-008-9032-0. Epub 2008 Feb 6.

Abstract

Previous work has shown that recombinant protein yield can be improved from in vitro-cultured mammalian cells by culturing at sub-physiological temperatures, although this effect is cell line and product dependent. The mechanism(s) by which low temperature leads to enhanced product yield are currently unknown; however, recent reports suggest that increased mRNA levels at sub-physiological temperatures may be largely responsible for this. Here, we have investigated whether low-temperature cultivation of cell lines selected for antibody production at 37 degrees C leads to changes in heavy- and light-chain mRNA levels and if this is reflected in antibody yields. Low-temperature in vitro culturing resulted in reduced viable cell concentration, prolonged cell viability, a reduction in metabolite consumption and production, cell cycle arrest in both CHO and NS0 cells, and changes in the levels of heavy- and light-chain mRNA. Despite increases in the level of heavy- and light-chain mRNA upon culturing at 32 degrees C in our model CHO cell line, this did not result in increased total product yield; however, changes in cell-specific yields were observed that reflected the metabolic rate of glucose utilization and changes in mRNA levels.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / genetics*
  • Antibodies, Monoclonal / metabolism*
  • Antibody Formation*
  • CHO Cells
  • Cell Cycle
  • Cell Proliferation
  • Cells, Cultured
  • Cricetinae
  • Cricetulus
  • Gene Expression Regulation
  • Glucose / metabolism
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Heavy Chains / metabolism
  • Immunoglobulin Light Chains / genetics
  • Immunoglobulin Light Chains / metabolism
  • Lactic Acid / metabolism
  • Mice
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Recombinant Proteins / biosynthesis
  • Temperature*

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Light Chains
  • RNA, Messenger
  • Recombinant Proteins
  • Lactic Acid
  • Glucose