Effect of osmolytes on the conformational stability of mouse monoclonal antidigoxin antibody in long-term storage

Hybridoma (Larchmt). 2008 Apr;27(2):99-106. doi: 10.1089/hyb.2007.0554.

Abstract

The natural selection of small organic molecules (osmolytes) that protect cellular proteins against denaturing stresses is characteristic of plants and animals that have adapted to environmental extremes. In this study the role of osmolytes on the structural stability of antibody in long-term storage was investigated. Native antibody preparations isolated from cell culture supernatant were formulated with different additives for storage stabilization. The additives studied were arginine, proline, and sorbitol. Structural stability of antibody investigated and effect of these osmolytes on retaining the native structure in long-term storage were established. CD spectra of native IgG showed the characteristics expected for beta-proteins. After the passing of given time, the beta-sheet structure of IgG was reduced but the addition of the osmolyte to antibody solution prevented loosing the ordered structures. These results can be effective for stabilization of proteins in commercial and industrial processes.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acids / physiology*
  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / metabolism
  • Arginine / physiology
  • Digoxin / immunology*
  • Mice
  • Osmotic Pressure / drug effects
  • Proline / physiology
  • Protein Conformation / drug effects
  • Protein Denaturation / drug effects
  • Protein Folding
  • Solutions
  • Sorbitol* / pharmacology
  • Spectrometry, Fluorescence
  • Thermodynamics
  • Time Factors

Substances

  • Amino Acids
  • Antibodies, Monoclonal
  • Solutions
  • Sorbitol
  • Digoxin
  • Arginine
  • Proline