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.
D000818:Animals; D000951:Antigens, Neoplasm / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000302:isolation & purification*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003840:Deoxycholic Acid; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification*; D008565:Membrane Proteins / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008811:Mice, Inbred DBA; D008970:Molecular Weight; D010954:Plasmacytoma / Q000276:immunology; D013154:Spleen / Q000166:cytology
D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D006023:Glycoproteins; D008565:Membrane Proteins; D003840:Deoxycholic Acid