United Kingdom Atomic Energy Authority

Patrick Mattock

Alan Rosevear

Gordon F Aitchison

Gordan F Aitchison

The extraction of Factor VIII, the antihaemophilic factor in blood, is difficult due to its instability and the presence of impurities. An initial Factor VIII containing aqueous solution such as blood plasma is purified by subjecting a Factor VIII containing aqueous migrant solution to continuous flow electrophoresis wherein flow takes place in an annular separation chamber and is stabilized by means of an angular velocity gradient; and collecting a separated Factor VIII component. In order to separate the Factor VIII from albumin and firbrinogen, while obtaining good recoveries of Factor VIII (e.g. .about.60%), the migrant solution is prepared by precipitating Factor VIII from the initial solution using ethanol, and removing and redissolving the precipitate in an aqueous medium and adjusting the pH to be within the range of 7.5 to 8.6, preferably 8.3 to 8.6.

Blood fractionation improvement

The invention discloses a process for the selective separation of a chemical component from a fluid substance which comprises contacting the fluid substance with a composite material having affinity chromatography properties, said composite material comprising an affinity chromatography agent retained within the pore structure of a porous rigid support material and said affinity chromatography agent being capable of selectively sorbing the chemical component from the fluid substance.

Affinity chromatography separation process

The invention discloses a composite material comprising a deformable gel retained within the pores of a porous rigid support material. The deformable gel comprises an affinity chromatography agent such that the composite material has affinity chromatography properties.

Composite materials comprising deformable xerogel within the pores of

PCT No. PCT/GB78/00038 Sec. 371 Date June 22, 1979 Sec. 102(e) Date June 22, 1979, PCT Filed Nov. 10, 1978 PCT Pub. No. WO 79/00299 PCT Pub. Date May 31, 1979. The invention is concerned with purification of Factor VIII containing solutions, such as blood plasma, by continuous flow electrophoresis. Hitherto, purification of such solutions has been performed by methods such as cryoprecipitation which however have the disadvantages of poor recovery. In our invention, this problem is overcome by adjusting the pH of a Factor VIII containing solution to be in a range where the stability of Factor VIII is not adversely affected (e.g. 6 to 9) and then subjecting the solution to continuous flow electrophoresis to give purified Factor VIII fractions. If desired, the fractions may be further purified.

Purification of Factor VIII

The invention is concerned with fractionation of proteins, in particular with fractionation of immunoglobulin containing solutions, such as blood plasma, by continuous flow electrophoresis. Hitherto, such solutions have been fractionated by ethanol precipitation methods (e.g. the Cohn Method). Such methods separate the IgG component of immunoglobulins but cannot separate the useful IgM component. In the invention, the solutions are fractionated by continuous flow electrophoresis at a pH of between 7 and 8.4 and an electrical conductivity of between 1 and 2 mScm.sup.-1, thereby giving rise to fractions containing particular combinations of immunoglobulin components (IgG, IgM, IgA, IgD and IgE) with potentially valuable properties.

Fractionation of proteins

Continuous flow electrophoresis in an apparatus such as described in U.S. Pat. No. 3,616,453 is stabilized by using a migrant solution containing a sufficient quantity of a liquid of density less than water such as ethanol. Continuous flow electrophoresis may be used, for example, in the fractionation and/or purification of a wide range of biologically active materials.

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Botley, United Kingdom

Patrick Mattock