Development of Large‐Scale Fractionation Methods
作者:
M. Wickerhauser,
J. E. Mercer,
J. W. Eckenrode,
期刊:
Vox Sanguinis
(WILEY Available online 1978)
卷期:
Volume 35,
issue 1‐2
页码: 18-31
ISSN:0042-9007
年代: 1978
DOI:10.1111/j.1423-0410.1978.tb02896.x
出版商: Blackwell Publishing Ltd
数据来源: WILEY
摘要:
Abstract.To increase purity and potency of the antihemophilic factor (AHF) concentrate prepared by the American Red Cross method, the following modifications were introduced: (1) A cold extraction step was incorporated to remove cold‐soluble impurities. The cryopre‐cipitate (cryo) was extracted with 0.02MTris buffer, pH 7.0 (4 ml/g cryo) at 0°C. Factor VIII loss in this step was negligible. (2) AHF was then recovered from the cold‐insoluble portion of the cryo by extraction at 21°C with the same buffer. To increase the AHF concentration, this second extraction step was carried out with a smaller buffer volume (2 ml instead of 3–4 ml/g cryo). The subsequent steps, deprothrombinization, filtration and lyophilization were essentially unchanged. To further increase factor VIII concentration, the dried AHF concentrate was reconstituted to 40 rather than 50% of the initial volume. AHF concentrate prepared on a large scale by this method was 20‐ to 30‐fold concentrated and 40‐ to 50‐fold purified over plasma at a recovery of about 250 factor VIII units per liter of plasma. The final product was readily soluble, clear and almost colorless up
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