Measurement of antigen-antibody binding constants by elution of affinity chromatography columns with continuous concentration gradients of dissociating agents.
Elution of antibody-Sepharose 4 B columns previously loaded with antigen by a linear concentration gradient of Na I or NaSCN, was used for the calculation of binding constants for the human IgG - anti human IgG system. The values obtained by this method showed good agreement with the values obtained by "stepwise" elution with different NaI concentrations, with the values obtained when the antigen was immobilized, and with the values determined in solution. The method employed appears as a simple and useful possibility for the determination or comparison of K values.
D000936:Antigen-Antibody Complex / Q000032:analysis*; D002846:Chromatography, Affinity / Q000379:methods*; D007700:Kinetics; D011485:Protein Binding; D012974:Sodium Iodide / Q000494:pharmacology; D013861:Thiocyanates / Q000494:pharmacology
D000936:Antigen-Antibody Complex; D013861:Thiocyanates; C024553:sodium thiocyanate; D012974:Sodium Iodide