HPLC analysis of the dissociation and recombination of rabbit immunoglobulin G.
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.
D000085:Acetates; D019342:Acetic Acid; D000818:Animals; D000918:Antibody Specificity; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004226:Dithioerythritol; D007074:Immunoglobulin G / Q000032:analysis*; D010084:Oxidation-Reduction; D011487:Protein Conformation; D011817:Rabbits
D000085:Acetates; D007074:Immunoglobulin G; D004226:Dithioerythritol; D019342:Acetic Acid