Partial purification of a membrane glycoprotein antigen by high-pressure size-exclusion chromatography without loss of antigenicity.

A high-pressure size exclusion chromatography system eluted with phosphate saline buffer containing 0.25% sodium deoxycholate has been developed that fractionates both soluble and membrane glycoproteins with good resolution and molecular weight versus elution time relationship. Using this system we fractionated membrane glycoproteins from a mutagenized mastocytoma cell that carries a strong transplantation antigen. After dialysis to remove the detergent, the fractions were tested for biological activity by an in vitro assay involving T-lymphocyte cell culture. Antigenic activity was found between 43 and 85 kdaltons. This demonstrates the efficiency of the system to resolve complex membrane protein samples without destroying their biological activity.