Determination of immunoglobulins in blood serum by high-performance affinity chromatography.
High-performance affinity chromatography columns were prepared by immobilizing protein A from Staphylococcus aureus on 10-microns diol-bonded silica. Immunoglobulin-containing samples were injected into the column at pH 7 and eluted by stepwise changes to pH 3. Immunoglobulin G, A, and M standards were tested, but only a small fraction of the latter two was retained on the column. Analysis of 3-microliters samples of reference blood sera without pretreatment was performed using immunoglobulin G standards. Good agreement was observed between the immunoglobulin G concentrations measured by this method and by radial immunodiffusion. Chromatography of albumin and electrophoresis of retained serum fractions showed no interference by non-specific adsorption. Chromatographic analysis times were 4 min or less.
D002846:Chromatography, Affinity; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007070:Immunoglobulin A / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000302:isolation & purification; D007075:Immunoglobulin M / Q000302:isolation & purification; D007136:Immunoglobulins / Q000302:isolation & purification*; D013205:Staphylococcal Protein A
D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007136:Immunoglobulins; D013205:Staphylococcal Protein A