Gel chromatography applied to quantitation of components of IgG preparations

Investigations were undertaken to standardize gel chromatography for quality control of IgG preparations. Great importance was attached to convenience of procedure. With application of the optimal type ofgel, Ultrogel AcA 34, the influence on quality of separation of various experimental parameters such as pH and ionic strength of the eluant, volume and concentration of the sample, flow rate and direction, and temperature were tested. Various methods of evaluation were applied. According to the demands, quantitation of components may be performed by collection of fractions and determination of peak volumes and protein concentrations, by determination of peak areas on absorbance recordings, or automatically by use of an integrator or a computer. The results obtained by gel chromatography are discussed and compared with the results of analytical ultracentrifugation.