Soluble HL-A antigens in serum. I. Isolation and Purification

✍ Scribed by S. K. Oh; M. A. Pellegrino; S. Ferrone; E. D. Sevier; R. A. Reisfeld

• Book ID: 102822393
• Publisher: John Wiley and Sons
• Year: 1975
• Tongue: English
• Weight: 693 KB
• Volume: 5
• Category: Article
• ISSN: 0014-2980
• DOI: 10.1002/eji.1830050302

✦ Synopsis

Abstract

A new method was developed which utilizes fractional salting out, gel filtration, ion exchange chromatography and polyacrylamide gel electrophoresis for the isolation and purification of soluble HL‐A antigens from serum. The method produced purified antigens possessing HL‐A 5,9 specificities with yield up to 60 % of that detected in the original serum. Up to 150‐fold increase in specific HL‐A antigenic activity above that in the starting material could be achieved as measured by the ability of the purified antigens to specifically inhibit the cytotoxic activity of HL‐A alloantisera against selected target cells. Electrophoretically purified HL‐A antigens with different specificities appear to have a similar molecular size, i.e. 33 000, but differ in their electric charge properties. β~2~‐microglobulin (β~2~m) seems to be noncovalently associated with HL‐A antigens present in serum, but during purification β~2~m is progressively lost until at the final purification by polyacrylamide gel electrophoresis, the material is completely devoid of β~2~m.