Novel application of H tube electrophoresis to separation technology.

Abstract

The H tube electrophoresis technique was used for isolating human IgG by the net effect of electrophoresis and electro-endosmosis in agarose gel containing human serum. In a similar experiment human serum albumin was purified by extracting the agarose gel slice containing the albumin following zone electrophoresis of serum in agarose. By taking advantage of the relatively high reverse electro-endosmotic flow compared to the migration of IgG in agarose gel, sheep anti chicken immunoglobulin was fractionated into active and inactive portions. It was estimated that 90% of the nonspecific IgG could be eliminated in this manner. The specific immunoglobulin fraction showed a higher mobility to the positive electrode than the bulk of the non specific IgG. The H tube proved to be ideal for zone electrophoresis in concentration gradients of inert substances. For this purpose sugar gradients were used for electrophoretic separation of proteins and viruses which are not affected by high osmotic pressures but, for separating animal cells it was necessary to use Ficoll gradients. A mixture of yellow Atypical Tomato Mosaic Virus and the haemocyanin of Burnupena cincta were separated into discrete zones and chicken IgY anti haemocyanin was subfractionated. The specific fraction showed a lower migration rate to the positive pole than the bulk of the non specific material.