HPLC analysis of the dissociation and recombination of rabbit immunoglobulin G

J Immunol Methods. 1987 Feb 11;96(2):195-9. doi: 10.1016/0022-1759(87)90314-0.

Abstract

Rabbit immunoglobulin G (RIgG) was reduced with dithioerythritol and analyzed by high performance liquid chromatography. A quantitative method for determining the percentage of reduced half-molecules in the mixture was developed. An acetic acid concentration-dependent rate of dissociation of reduced half-molecules was observed. The specific optical absorptivity was determined for whole molecules and half-molecules and found to be significantly greater for the half-molecules. Purified half-molecules were reconstituted into RIgG with a yield greater than 90% following a 16 h incubation at pH 8.0 and room temperature.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetates
  • Acetic Acid
  • Animals
  • Antibody Specificity
  • Chromatography, High Pressure Liquid / methods*
  • Dithioerythritol
  • Immunoglobulin G / analysis*
  • Oxidation-Reduction
  • Protein Conformation
  • Rabbits

Substances

  • Acetates
  • Immunoglobulin G
  • Dithioerythritol
  • Acetic Acid