Large-Scale Gel Filtration Chromatography for the Production of a Solvent/Detergent-Treated High-Purity Factor VIII Concentrate

Vox Sanguinis ◽  
1990 ◽  
Vol 58 (4) ◽  
pp. 257-263
Author(s):  
U. Stöcker ◽  
T. Dengler ◽  
G. Fürst ◽  
S. Keller
Keyword(s):  
Factor Viii ◽  
High Purity ◽  
Large Scale ◽  
Gel Filtration ◽  
Gel Filtration Chromatography

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.

scholarly journals Conditions Contributing to the Stability of Factor VIII Coagulant Activity

1977 ◽  
Author(s):  
T. Exner ◽  
K.A. Rickard ◽  
H. Kronenberg
Keyword(s):  
Factor Viii ◽  
High Purity ◽  
Gel Filtration ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Buffer System ◽  
Factor Viii Activity ◽  
Coagulant Activity ◽  
The Stability ◽  
Degree Of Purification

Factor VTII tends to become less stable the greater its degree of purification. The loss of factor VIII during preparation of high activity concentrates makes such processes uneconomical. Conditions contributing to the stability of factor VIII were investigated.High purity factor VIII was incubated with plasma components fractionated by gel filtration and by anion exchange chromatography. Factor VIII activity was assessed initially and after several hours incubation. Several fractions destroying factor VIII activity were clearly resolved. Fractions stabilizing factor VIII were associated only with albumin.Various buffer systems were investigated similarly. A non-chelating buffer system containing albumin was found to give optimal factor VIII stability.

Large-scale isolation of covalently closed circular DNA using gel filtration chromatography

1988 ◽  
Vol 173 (1) ◽  
pp. 125-133 ◽  
Author(s):  
Gregory J. Raymond ◽  
P.Kruger Bryant ◽  
Arlen Nelson ◽  
Jerry D. Johnson
Keyword(s):  
Large Scale ◽  
Gel Filtration ◽  
Gel Filtration Chromatography ◽  
Circular Dna ◽  
Covalently Closed Circular Dna

scholarly journals Large Scale Purification and Characterisation of Porcine Factor VIII for Clinical Use

1977 ◽  
Author(s):  
Helen Lee ◽  
S.I. Chavin ◽  
Rita E. Blewitt ◽  
D. E. G. Austen
Keyword(s):  
Factor Viii ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Large Scale ◽  
Gel Filtration ◽  
Cross Reactivity ◽  
Value Analysis ◽  
Human Red Blood Cells ◽  
Human Factor Viii ◽  
Haemagglutinating Activity

Animal factor VIII can be useful in the treatment of haemophilic patients with high titres of human factor VIII antibody when the antibody shows low cross-reactivity towards animal factor VIII. The use of currently available preparations of animal factor VIII is associated with rigor, anaphylaxis and transient thrombocytopaenia, which limit their therapeutic value. Analysis of some of these preparations has shown low IgM and high IgG concentrations and haemagglutinating activity against a panel of human red blood cells. It has been demonstrated that purified porcine IgG and IgN agglutinate human red blood cells, while IgM is also complement fixing.A large scale purification procedure has been developed for producing porcine factor VIII from platelet-poor plasma. It involves various precipitations followed by gel filtration through Sepharose 4B-CL on a KS sectional column. The pooled void-volume fractions are concentrated by using the Pellicon cassette system, giving a final yield of approximately 20% of the starting plasma. Porcine factor VIII prepared by this method is stable. Its greatly reduced fibrinogen content facilitates final filtration and prevents the accumulation of fibrinogen in patients treated with high doses. Non-specific haemagglutinins which may have contributed to some of the post-transfusion reactions have been removed. Specific activities of at least 50 U/mg of protein are obtained, and doses greater than 10,000 units can be given in 10 ml.

scholarly journals Purification and heterogeneity of inorganic pyrophosphatase of pig scapula cartilage

1975 ◽  
Vol 147 (1) ◽  
pp. 103-109 ◽  
Author(s):  
R Felix ◽  
H Fleisch
Keyword(s):  
Enzyme Activity ◽  
Ion Exchange ◽  
High Purity ◽  
Cetylpyridinium Chloride ◽  
Gel Filtration ◽  
Inorganic Pyrophosphatase ◽  
Gel Filtration Chromatography ◽  
Enzyme Preparations ◽  
Specific Activities

Inorganic pyrophosphatase (pyrophosphate phosphohydrolase; EC 3.6.1.1) was purified from pig scapula cartilage by fractionation with N-cetylpyridinium chloride and (NH4)2SO4, followed by ion-exchange and gel-filtration chromatography. Enzyme preparations of high purity were obtained, with specific activities (100-400 units/mg) higher than those previously described for mammalian pyrophosphatases. The enzyme activity could be separated into several subfractions on ion-exchange columns.

Importance of Protease Inhibition in Studies on Purified Factor VIII (Antihaemophilic Factor)

1976 ◽  
Vol 35 (01) ◽  
pp. 186-190 ◽  
Author(s):  
Eugen A. Beck ◽  
Peter Bachmann ◽  
Peter Barbier ◽  
Miha Furlan
Keyword(s):  
Ionic Strength ◽  
Factor Viii ◽  
Gel Filtration ◽  
High Ionic Strength ◽  
Procoagulant Activity ◽  
Carrier Protein ◽  
Protease Inhibition ◽  
Functional Sites ◽  
Molecular Weight Peak ◽  
Von Willebrand

SummaryAccording to some authors factor VIII procoagulant activity may be dissociable from carrier protein (MW~ 2 × 106) by agarose gel filtration, e.g. at high ionic strength. We were able to reproduce this phenomenon. However, addition of protease inhibitor (Trasylol) prevented the appearance of low molecular weight peak of factor VIII procoagulant activity both at high ionic strength and elevated temperature (37°C). We conclude from our results that procoagulant activity and carrier protein (von Willebrand factor, factor VIII antigen) are closely associated functional sites of native factor VIII macro molecule. Consequently, proteolytic degradation should be avoided in functional and structural studies on factor VIII and especially in preparing factor VIII concentrate for therapeutic use.

Factor VIII Concentrate: What is “High Purity”?

1994 ◽  
Vol 72 (03) ◽  
pp. 483-484 ◽  
Author(s):  
Axel Schoppmann ◽  
Alfred Weber ◽  
Felix Hondl ◽  
Yendra Linnau
Keyword(s):  
Factor Viii ◽  
High Purity
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