Abstract 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. ˜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.
In a method of purifying an initial Factor VIII containing aqueous solution which comprises subjecting a Factor VIII containing aqueous migrant solution to continuous flow electrophoresis by injecting the solution into a second aqueous solution, laminarly flowing in an annular separation chamber as a carrier solution for the migrant solution and stabilised by means of an angular velocity gradient, and applying a constant electric field across the resulting mixture to produce a differential movement of the Factor VIII component of the migrant solution with respect to the other major components of the solution perpendicular to the direction of flow of the layer; and collecting the separated Factor VIII component, the improvement wherein the migrant solution is prepared by (i) contacting the initial Factor VIII containing aqueous solution with ethanol under conditions to precipitate Factor VIII; and
Claim 2
Dependent Claim
136 words
In a method of purifying an initial Factor VIII containing aqueous solution which comprises subjecting a Factor VIII containing aqueous migrant solution to continuous flow electrophoresis by injecting the solution into a second aqueous solution, laminarly flowing in an annular separation chamber as a carrier solution for the migrant solution and stabilised by means of an angular velocity gradient, and applying a constant electric field across the resulting mixture to produce a differential movement of the Factor VIII component of the migrant solution with respect to the other major components of the solution perpendicular to the direction of flow of the layer; and collecting the separated Factor VIII component, the improvement wherein the migrant solution is prepared by (i) contacting the initial Factor VIII containing aqueous solution with ethanol under conditions to precipitate Factor VIII; and
Claim 3
Dependent Claim
23 words
(ii) removing and redissolving the precipitation in an aqueous medium and adjusting the pH to be within the range of 7.5 to 8.6.
Claim 4
Dependent Claim
23 words
(ii) removing and redissolving the precipitation in an aqueous medium and adjusting the pH to be within the range of 7.5 to 8.6.
Claim 5
Dependent Claim
22 words
A method as claimed in claim 1 wherein, the step (ii), the pH is adjusted to a pH between 8.3 and 8.6.
Claim 6
Dependent Claim
21 words
A method as claimed in claim 1 wherein the initial Factor VIII containing solution is in the form of blood plasma.