Use of monoclonal antibodies for weak affinity chromatography.
Weak monoclonal antibodies were used as ligands in a high-performance liquid affinity chromatography system. Isocratic weak affinity chromatography was achieved when similar carbohydrate antigens were separated according to their weak binding to the immobilized monoclonal antibody. These chromatographic systems were studied in detail in terms of affinity, specificity and efficiency. The influence of the physico-chemical factors of temperature, pH, ionic strength and organic solvents was also evaluated. The issue of specificity was specially considered, as non-specific interactions are prevalent and usually of a weak affinity nature. This study clearly demonstrates the potential to use weak affinity biological interactions as the basis of chromatographic analysis and separation.
Ohlson
S
S
University of Kalmar, Department of Natural Sciences, Sweden.
Bergström
M
M
Påhlsson
P
P
Lundblad
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000941:Antigens / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D002241:Carbohydrates / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods*; D006825:Hybridomas; D006863:Hydrogen-Ion Concentration; D051379:Mice; D009994:Osmolar Concentration; D012680:Sensitivity and Specificity; D012997:Solvents / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
D000911:Antibodies, Monoclonal; D000941:Antigens; D002241:Carbohydrates; D012997:Solvents