In the blood of humans and animals, various substances participating in the coagulation of blood (blood coagulation factors) are contained. Various disorders or diseases induced from the shortage or lack of blood coagulation factors have been known. In particular, hemophilia A, due to the lack of blood coagulation factor VIII, occupies almost 80% of the whole of such diseases. There has hitherto been proposed no effective therapy for a radical cure of hemophilia A and, up to date, a therapeutic measure of replenishing the patient with blood coagulation factor VIII on requirement has currently been employed.
Blood coagulation factor VIII consists of a protein having a molecular weight of about 260,000, which is contained in the normal blood plasma only in an amount of about 0.1 .mu.g/ml. Blood coagulation factor VIII is present in blood plasma under formation of a complex with another blood coagulation factor, namely, von Willebrand factor (abbreviated hereinafter as vWf) composed of proteins having molecular weights of 500,000-20,000,000. In the context of this application, by the word "blood coagulation factor" is meant to include such a complex. In the therapy of hemophilia A, no sufficient replenishment of blood coagulation factor VIII can be achieved by transfusion of an intact whole blood or plasma as such to the patient, since the concentration of blood coagulation factor VIII in the plasma is quite low. It has therefore been a wide spread practice to employ an intravenous injection of a concentrated preparation of blood coagulation factor VIII for such a patient.
It has been practiced to carry out concentration of blood coagulation factor VIII by forming first a cryoprecipitate from blood plasma, separating it and then thawing it to obtain a concentrate, or by employing purification by liquid chromatography and so on. However, the yield of recovery of blood coagulation factor VIII in these prior techniques has been able to reach to a figure of only about a little lower than 10%, based on the weight of the plasma, and even by the technique using a liquid chromatography, to a value of about 20%.
In the technique employing liquid chromatography, separation materials have been employed, in which ligands, such as, substances having affinity to blood coagulation factor VIII or to vWf, for example, substances having aminoalkyl groups (Japanese Patent Kokai Nos. 136518/1985 and 157000/1989), monoclonal antibodies (Japanese Patent Kokai Nos. 13099/1989 and 221396/1989), collagens (Japanese Patent Kokai No. 198632/1988 and so on) and so on are caused to link onto the substrate.
For a separation material for use for recovering blood coagulation factor VIII in a liquid chromatography, the following properties are required:
1) It should not cause any non-specific adsorption of plasma proteins. PA0 2) It should have a pore diameter capable of permeating a complex of blood coagulation factor VIII with vWf and a sufficiently wide internal surface area for making avail of a large adsorption capacity. PA0 3) The substrate to be combined with the ligand should have high stabilities not only in physical but also in chemical properties. PA0 (1) A separation material for separating and recovering a blood coagulation factor, comprises a porous matrix having linked thereon one or more ligands each consisting of a radical exhibiting an affinity to the blood coagulation factor to be recovered, said porous matrix having a specific surface area of at least 1.5 m.sup.2 per milliliter of separation material with respect to pores having diameters of at least 0.1 .mu.m and said porous matrix being derived from a porous particulate material having an exclusion limit molecular weight of at least 1.5.times.10.sup.6 as determined with polyethylene glycol. PA0 (2) A process for preparing the separation material for separating and recovering a blood coagulation factor, comprising the steps of PA0 (3) A process for recovering a blood coagulation factor from a blood coagulation factor-containing raw material using a separation material which comprises a porous matrix having linked thereon one or more ligands each consisting of a radical exhibiting an affinity to the blood coagulation factor to be recovered, said matrix having a specific surface area of at least 1.5 m.sup.2 per milliliter of the separation material with respect to pores having diameters of at least 0.1 .mu.m and said matrix being derived from a porous particulate material having an exclusion limit molecular weight of at least 1.5.times.10.sup.6 as determined with polyethylene glycol, said process comprizing the steps of
In the conventional techniques mentioned above, separation materials prepared from natural high molecular weight substances, such as, agarose, dextran, cellulose and so on are employed. These separation materials are superior in so far as they employ substrates consisting of natural substances and non-specific adsorption will scarcely occur. For these prior art separation material no quantitative assay as to the properties mentioned in the above 2) has been made, so that it might have hitherto been probable that separation material having lower adsorption capacity are employed with poor recovery yield of blood coagulation factors.