Isolation and radioimmunoassay of human transplantation antigens

✍ Scribed by N. Tanigaki; Y. Miyakawa; V. P. Kreiter; D. Pressman

• Publisher: John Wiley and Sons
• Year: 1972
• Tongue: English
• Weight: 719 KB
• Volume: 4
• Category: Article
• ISSN: 0022-4790
• DOI: 10.1002/jso.2930040209

✦ Synopsis

A procedure has been developed for the isolation of relatively pure human transplantation antigens in high yield from established cultures of human hematopoietic cells. Several preparations of soluble cell fragments, each carrying as the evident activity one of the specificities, HL-Al, HL-A2, and HL-A7, have been isolated in a state of high molecular purity and high specific activity. Cell particulates, obtained by the disruption of cells when they were agitated in buffer, were digested under carefully controlled conditions with papain in the presence of cysteine. The soluble fiagments obt+ined were carried through a series of purification steps-hypotonic dialysis, DEAE ion exchange chromatography, ultrafiltration, gel filtration, and column electrophoresis. The final preparations contained 30-37% of the total HL-A activity present in the papain digest. All fragments carrying HL-A activities had a molecular size of about 48,000 Daltons. Some of the HL-A specificities were associated with differences in electrophoretic mobility of the fragments, and, on this basis, different specificites found in a single cell line could sometimes be separated from each other. The products were labeled with radioiodine without affecting the antigenic activity. These labeled antigens were used for a number of purposes. The chief application has been as a reagent in a sensitive radioimmunoassay method for HL-A activity that is applicable to whole cells or particulates as well as to soluble antigens.

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