Effect of Gelatin on the Stability of Heavy Chain Monoclonal Antibody Production from Plant Suspension Cultures

  • Ryland, J. (Washington State University) ;
  • Robert, P. (Washington State University) ;
  • Michael, Linzmaier (Washington State University) ;
  • Lee, James M. (Washington State University)
  • Published : 2000.08.01

Abstract

The heavy chain monoclonal antibody (HC MAb) was produced in suspension cultures of genetically modified Nicotiana tabacum. The HC MAb secreted to the medium was unstable due to unfavorable interactions in the plant cell medium. The addition of gelatin (5g/l) stabilized the extracellular HC MAb and increased its production 10-fold. A kinetic model was developed describing the interaction between the secretedprotein and the stabilizer. The model accounted for the inactivation of the protein by simple aggregation and general instability. It was assumed that the secreted protein and the stabilizer form a stable complex. Culturing the cells semicontinuously could further increase the productivity of HC MAb.

Keywords

References

  1. Science v.260 Chaperones: Helpers aling the pathway to protein folding Craig, E. A.
  2. Antibodies: A Laboratory Manual Harlow, E.;D. Lane.
  3. Proc. Nat. Acad. Sci. USA v.87 High level production of a functional immunoglobulin heterodimer in a baculovirus expression system Hasemann, C.;J. Capra
  4. J. Cell Biol. v.132 Roles of molecular chaperones in protein degradation Hayes, S. A.;J. F. Dice
  5. J. Phys. Chem. v.94 Measurement of the interaction between absorbed polyeletrolytes: Gelatin on mica surfaces Kawashini, N,;H, Christenson;B. Niham
  6. Biotechnol. Lett. v.19 The effect of polyvinylpyrrolidone (PVP) on the heavy chain monoclonal antibody production from plant suspension cultures LaCount, W.;G. An;J. M. Lee.
  7. Plant Physiol. v.82 Design and testing of a GRG code in liquid culture Lasdon, L. S.;A. D. Waren;A. Jain;M. Ratner
  8. J. Protein Exp. Purification v.7 Enhanced recovery of a secreted mammalian protein from suspension culture of genetically modified tobacco cells Magnuson, N. S.;P. M. Linzmaier;J. Gao;R. Reeves;G. An;J. M. Lee
  9. Plant Cell Rep v.15 A simple, accurate method for determining wet and dry weight concentrations of plant cell suspension cultures Mills, D.;J. M. Lee
  10. Physiol. Plant v.15 A revised medium for rapid growth bioassays with tobacco tissue cultures Murashige, T.;F. Skoog
  11. Comp. J. v.7 A simplex method for functional minimization Nelder, J. A.;R. Mead
  12. The Macromolecular Chemistry of Gelatin Veis, A.
  13. Biotechnol. Lett. v.17 Effects of dimethyl sulfoxide on heavy chain monoclonal antibody production from plant cell culture Wahl, M. F.;J. M. Lee
  14. Macromolecules v.26 Simultaneous calibration of size exclusion chromatography and dynamic light scattering for the characterization of gelatin Wu, C.