scholarly journals An Improved Method for Preparation of Antihemophilic Factor (AHF)

1977 ◽  
Author(s):  
M. Wickerhauser ◽  
J.E. Mercer ◽  
J.W. Eckenrode
Keyword(s):  
Factor Viii ◽  
Large Scale ◽  
Original Method ◽  
Home Treatment ◽  
Tris Buffer ◽  
Red Cross ◽  
American Red Cross ◽  
Improved Method ◽  
Extraction Step ◽  
Antihemophilic Factor

Intermediate purity AHF prepared by the American Red Cross method (James, H. L. and Wicker-hauser, M., Vox Sang. 23:402, 1972) contains about 10 Factor VIII units/ml at 20-25 fold purification over plasma. A more concentrated and purified product would facilitate administration of AHF in home treatment of hemophiliacs. Our original method was improved by the following modifications: (1) A cold extraction step was incorporated to remove cold-soluble impurities. The cryoprecipitate was extracted by stirring with 0°02 M Tris buffer, pH 7°0 (4 ml/g cryo) for 30 minutes at 0°CoFactor VIII loss in this step was negligible,,(2) AHF was then recovered from the cold-insoluble portion of the cryoprecipitate 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 subseguent steps, deprothrombinazation, filtration, and lyophi1ization were essentially unchanged,, To further increase Factor VIII concentration, the dried AHF concentrate was reconstituted to 40% rather than 50% of the volume prior to lyophilization. AHF concentrate prepared on a large scale by this method was 20-30 fold concentrated and 40-50 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 upon reconstitution.

Purification of the Antihemophilic Factor by Gel Filtration on Agarose

1969 ◽  
Vol 22 (03) ◽  
pp. 577-583 ◽  
Author(s):  
M.M.P Paulssen ◽  
A.C.M.G.B Wouterlood ◽  
H.L.M.A Scheffers
Keyword(s):  
Factor Viii ◽  
Plasma Proteins ◽  
Large Scale ◽  
Gel Filtration ◽  
Large Scale Preparation ◽  
Scale Preparation ◽  
Antihemophilic Factor

SummaryFactor VIII can be isolated from plasma proteins, including fibrinogen by chromatography on agarose. The best results were obtained with Sepharose 6B. Large scale preparation is also possible when cryoprecipitate is separated by chromatography. In most fractions containing factor VIII a turbidity is observed which may be due to the presence of chylomicrons.The purified factor VIII was active in vivo as well as in vitro.

SPECIAL REVIEWS

PEDIATRICS ◽  
1948 ◽  
Vol 2 (4) ◽  
pp. 489-497
Author(s):  
CHARLES A. JANEWAY
Keyword(s):  
Plasma Proteins ◽  
Chemical Structure ◽  
Large Scale ◽  
Blood Donors ◽  
Red Cross ◽  
American Red Cross ◽  
Scale Separation ◽  
Medical Progress ◽  
Scientific Curiosity ◽  
Free People

This brief review of some of the recent accessions to our knowledge of the chemical structure, physiologic functions, and therapeutic applications of the plasma proteins serves to emphasize three important elements in medical progress—scientific curiosity, the humanitarian impulse, and effective social organization. We have had the privilege of summarizing the work of hundreds of investigators, whose studies are giving us new tools for the investigation and treatment of disease. Their work has only been possible because the magnificent response of a free people to the call for blood donors by a voluntary philanthropic agency, the American Red Cross, was coupled with a technical triumph, the development of practical methods for the large-scale separation of the plasma proteins, itself the culmination of highly theoretical and seemingly impractical investigations by protein chemists in various countries for many years.

scholarly journals Factor VIII Concentrates - Forces of Supply and Demand

1977 ◽  
Author(s):  
Anthony F. H. Britten
Keyword(s):  
Factor Viii ◽  
Supply And Demand ◽  
The United States ◽  
Red Cross ◽  
American Red Cross ◽  
Treatment Demand ◽  
Choice Of Treatment ◽  
Factor Viii Concentrates ◽  
Total Freedom ◽  
Increasing Demand

Some surprisingly simple artificial factors influence the availability of factor VIII for hemophilia treatment. Demand is affected by choice of product, and choice of treatment style (episodic vs. prophylactic), and also by availability funds.Autobiographic data suggest the feasibility of long-term low dosage prophylaxis (6u/KgA8 hrs.). This tentative conclusion is based on 2½yrs. of unprecedented near-total freedom from hemorrhage. The significance is the potential saving of 60% of factor VIII concentrate, compared with the more orthodox dose (15u/KgA8 hrs.) which has been shown to increase factor VIII usage by 2.5–3 times compared with episodic therapy.The American Red Cross produced 293,332 units of cryoprecipitate from 4,690,217 whole blood donations in 1975-6 (6.25% of blood collected). Production could readily be increased if the demand existed. The following artificial factors have an influence on cryoprecipitate production: health insurance policies; price structure; the international market for factor VIII.It is concluded that there is no shortage of available factor VIII, that more could readily be made available, that prophylaxis may be possible without increasing demand for factor VIM, that financial considerations are seriously distorting hemophi1ia treatment, that hemophilia treatment in the United States is being subsidised by other countries, and that understanding of these factors can help to overcome them.

The Recovery of Factor VIII from Fresh-Frozen, Indated and Outdated Human Plasma

1976 ◽  
Vol 36 (01) ◽  
pp. 071-077 ◽  
Author(s):  
Daniel E. Whitman ◽  
Mary Ellen Switzer ◽  
Patrick A. McKee
Keyword(s):  
Factor Viii ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
The United States ◽  
Fresh Frozen Plasma ◽  
Red Cross ◽  
Random Samples ◽  
Fresh Frozen ◽  
Factor Viii Concentrates ◽  
Frozen Plasma

SummaryThe availability of factor VIII concentrates is frequently a limitation in the management of classical hemophilia. Such concentrates are prepared from fresh or fresh-frozen plasma. A significant volume of plasma in the United States becomes “indated”, i. e., in contact with red blood cells for 24 hours at 4°, and is therefore not used to prepare factor VIII concentrates. To evaluate this possible resource, partially purified factor VIII was prepared from random samples of fresh-frozen, indated and outdated plasma. The yield of factor VIII protein and procoagulant activity from indated plasma was about the same as that from fresh-frozen plasma. The yield from outdated plasma was substantially less. After further purification, factor VIII from the three sources gave a single subunit band when reduced and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. These results indicate that the approximately 287,000 liters of indated plasma processed annually by the American National Red Cross (ANRC) could be used to prepare factor VIII concentrates of good quality. This resource alone could quadruple the supply of factor VIII available for therapy.

Sign in / Sign up

Export Citation Format

Share Document