PROCEDURE FOR THE PREPARATION OF PURE INTRINSIC FACTOR AND OR NON-INTRINSIC FABTOR FROM PREPARATIONS CONTAINING BOTH SUBSTANCES

摘要

1327565 Intrinsic factor MEDIX AB OY 8 March 1972 [9 March 1971] 10748/72 Heading A5B Pure intrinsic factor and/or non-intrinsic factor may be isolated from a preparation containing both by (a) immunizing an animal with a material containing a vitamin-B12-binding macromolecule of the R1-type but lacking intrinsic factor, (b) adding an antiserum or antibody-containing immunoglobulin from the animal to the preparation containing intrinsic factor and non-intrinsic factor in such a quantity that all the non-intrinsic factor is bound to the antibody and (c) separating the intrinsic factor from the non-intrinsic factor/ antibody complex. The material containing a vitamin-B12-binding macromolecule may be a leukocyte-rich preparation obtained by centrifuging pig's blood, or may be saliva, bite, serum or cerebrospinal fluid or purified R-type vitamin-B12-binding substance. When an antibody containing immunoglobulin is used in step (b), it may be coupled with a solid carrier or another macromolecular material. The intrinsic factor and the non-intrinsic factor/antibody complex may be separated by electrophoresis, isoelectric focussing, gel filtration, ionexchange chromatography, or precipitation with salts, organic solvents or by an antigen-antibody reaction. Non-intrinsic factor may be isolated by dissociation of the antibody complex.