Comparison of non-biospecific effects in immunoaffinity chromatography using cyanogen bromide and bifunctional oxirane as immobilising agents
Murphy, R.F.; Conlon, J.M.; Imam, A.; Kelly, G.J.
Journal of Chromatography 135(2): 427-433
1977
ISSN/ISBN: 0021-9673 PMID: 874026 Document Number: 112119
Polypeptide antigen, glucagon, antibodies to glucagon and non-immune globulins were immobilized on agarose using CNBr and a bifunctional oxirane. Irrespective of the ligand immobilized, positively charged groups introduced to conjugates by CNBr caused electrostatic interactions with impurities and soluble biospecific ligands. Solvents required for elution of bound antibodies and antigens were more strongly deforming when immunoaffinity conjugates were prepared with CNBr than with the oxirane. This is attributed to compound affinity resulting from reinforcement of biospecific by non-biospecific interactions. Strongly deforming solvents were still required for oxirane conjugates when antibodies had high affinity for antigen.