Purification of human fibrinopeptides by high-performance liquid chromatography

✍ Scribed by James A. Koehn; Robert E. Canfield

• Publisher: Elsevier Science
• Year: 1981
• Tongue: English
• Weight: 601 KB
• Volume: 116
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(81)90370-5

✦ Synopsis

A simple two-step procedure for obtaining highly purifed human tibrinopeptides is described. This technique utilizes batch adsorption to a Clll silica-coated matrix packed in a disposable cartridge. This step serves both to concentrate and desalt fibrinopeptides released into the supernatant during the clotting of human fibrinogen. These peptides can be eluted from the cartridge in high yield with 50% acetonitrile and further purified by reverse-phase high-performance liquid chromatography on a FBondapak Cls column. Preparations of highly purified peptides have been obtained in yields greater than 80%. During the course of these studies, it became apparent that proteolytic degradation of tibrinopeptide B to des-Arg-FPB (Bfll-13) can substantially reduce the yields of fibrinopeptide B. Steps to prevent this reaction from occurring and a technique to isolate this degradation product are described.