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DOI: 10.1055/s-0038-1661438
Heparin Cofactor II: Purification and Antibody Production
Publikationsverlauf
Received 08. Juli 1985
Accepted 09. Oktober 1985
Publikationsdatum:
19. Juli 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/198601/lookinside/thumbnails/10-1055-s-0038-1661438-1.jpg)
Heparin cofactor II (HCII) was purified from plasma to homogeneity. The procedure includes adsorption with (Al)OH3, fractionation with polyethylene glycol 6000, chromatography on QAE-Sephadex A-50, on heparin-Sepharose 4B and on Sephadex G-150. QAE-Sephadex A-50 chromatography provides a good separation of HCII from antithrombin III (AT) and most contaminants having a heparin affinity similar to that of HCII. HCII is eluted at 0.28 M NaCl from the heparin-Sepharose column. After gel filtration on G-150, contaminating AT was removed by immunoadsorption. Purified HCII shows an apparent Mr of 66,500 dal tons as analyzed on SDS-polyacrylamide gel and 62,100 daltons by ultracentrifugation. Antibodies to HCII were raised in rabbits. Former antisera mostly directed to a contaminating protein were used to remove it from the HCII preparation. Antibodies to HCII were made monospecific by immunoadsorption on HCII-free plasma linked to Sepharose 4B. Since many functional AT assays have neglected the presence of HCII in plasma, antibodies to HCII using as immunoadsorbent will provide a more specific test for AT.
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