Document Detail


Studies on the purification and characterization of human factor 8.
MedLine Citation:
PMID:  4626584     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Factor VIII (antihemophilic globulin) has been prepared from Hyland method IV AHG and cryoprecipitate using limited chymotryptic digestion followed by Sepharose gel filtration. The activity of factor VIII is unaffected by the digestion procedure, while fibrinogen in converted to large noncoagulable fragments. The purified factor VIII has been found to be a macromolecular glycoprotein with a major subunit of 240,000, as shown by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Carbohydrate analysis of factor VIII gave values of 1% sialic acid, 2.8% hexosamine, and 1-2% hexose (mannose, galactose, and fucose). The lipid content was found to be less than 5% of the protein content, and included no detectable phospholipid. The amino acid content is also reported. Immunoelectrophoretic analysis using rabbit antibody to purified factor VIII produced a single precipitin line. The chymotrypsin digestion step facilitates the preparation of factor VIII by reducing the viscosity of fibrinogen in the crude starting material, thereby increasing fivefold the quantity of material which can be processed at one time. It also improves markedly the resolution between factor VIII and fibrinogen on gel filtration.
Authors:
S L Marchesi; N R Shulman; H R Gralnick
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Journal of clinical investigation     Volume:  51     ISSN:  0021-9738     ISO Abbreviation:  J. Clin. Invest.     Publication Date:  1972 Aug 
Date Detail:
Created Date:  1972-10-18     Completed Date:  1972-10-18     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7802877     Medline TA:  J Clin Invest     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2151-61     Citation Subset:  AIM; IM    
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / analysis
Carbamates
Chromatography, Gel
Chymotrypsin
Cold Temperature
Electrophoresis, Disc
Factor VIII / analysis,  isolation & purification*
Fibrinogen
Hexoses / analysis
Humans
Immunodiffusion
Immunoelectrophoresis
Lipids / analysis
Methods
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Carbamates; 0/Hexoses; 0/Lipids; 9001-27-8/Factor VIII; 9001-32-5/Fibrinogen; EC 3.4.21.1/Chymotrypsin
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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