Separation of serum proteins by high performance liquid chromatography

ABSTRACT

This article describes the separation of serum proteins by isocratic HPLC using a polyvinyl alcohol gel as the stationary phase. The system separates immunoglobulins and other proteins by size exclusion and weak affinity bonding and gives a chromatogram that can be evaluated qualitatively, in a manner similar to serum protein electrophoresis scans. Peak heights measured for IgG, IgA, and IgM correlate closely with concentrations measured by radial immunodiffusion (RID). The separation, carried out without sample pretreatment, may prove to be a valuable technique for quantifying several serum proteins. The method may be used to screen hybridoma cultures for antibody production and to purify proteins without risk of denaturation.