Preparation of α-L antibody, purification of α-L antigen and reagent for detection of α-L antibody and α-L antigen

Claim 1

A process for the preparation of an α-L antibody which comprises immunizing an animal, other than a human, with an α-L antigen or an α-L antigen/α-L antibody complex and collecting antiserum from the animal, said α-L antigens having the following properties: (1) Molecular weight: About 1,000,000; (2) Electrophoretic mobility: In the β--α 2 globulin region, which is an intermediate value between that of an HBs antigen and an α-fetoprotein; (3) Isoelectric point: PI = 5.04, the activity of the α-L antigen was shown in the range of 4.22 to 5.68 by measuring with an Ampholien column; (4) Buoyant Density: d = 1.08 g/ml in a solution of cesium chloride; and d = 1.03 g/ml in a solution of sucrose; (5) Formation of complex: A precipitable complex is formed with dextran sulfate in the presence of magnesium chloride, exhibiting lipoprotein-like properties; (6) Staining: The α-L antigen/antibody precipitated line in agar gel can be stained with Sudan black B; (7) thermostability: The antigenic properties in the serum are maintained at a temperature of 56° C. for 30 minutes; and (8) Form (as an antigen/antibody complex): the spherical particles are about 300 A in diameter.

Claim 12

A method for the purification of an α-L antigen which comprises preparing a solution of an α-L antigen and subjecting said solution to a density gradient centrifugation to collect a fraction having a buoyant density less than 1.1 g/ml, said α-L antigen having the following properties: (1) Molecular weight: About 1,000,000; (2) Electrophoretic mobility: In the β--α 2 globulin region, which is an intermediate value between that of an HBs antigen and an α-fetoprotein; (3) Isoelectric point: PI = 5.04, the activity of the α-L antigen was shown in the range of 4.22 to 5.68 by measuring with an Ampholien column; (4) Bouyant Density: d = 1.08 g/ml in a solution of cesium chloride; and d = 1.03 g/ml in a solution of sucrose; (5) Formation of complex: A precipitable complex is formed with dextran sulfate in the presence of magnesium chloride, exhibiting lipoprotein-like properties; (6) Staining: The α-L antigen/antibody precipitated line in agar gel can be stained with Sudan black B; (7) thermostability: The antigenic properties in the serum are maintained at a temperature of 56° C. for 30 minutes; and (8) Form (as an antigen/antibody complex): the spherical particles are about 300 A in diameter.

Claim 13

A method for the purification of an α-L antigen which comprises preparing a solution of an α-L antigen and subjecting said solution to a density gradient centrifugation to collect a fraction having a buoyant density less than 1.1 g/ml, said α-L antigen having the following properties: (1) Molecular weight: About 1,000,000; (2) Electrophoretic mobility: In the β--α 2 globulin region, which is an intermediate value between that of an HBs antigen and an α-fetoprotein; (3) Isoelectric point: PI = 5.04, the activity of the α-L antigen was shown in the range of 4.22 to 5.68 by measuring with an Ampholien column; (4) Bouyant Density: d = 1.08 g/ml in a solution of cesium chloride; and d = 1.03 g/ml in a solution of sucrose; (5) Formation of complex: A precipitable complex is formed with dextran sulfate in the presence of magnesium chloride, exhibiting lipoprotein-like properties; (6) Staining: The α-L antigen/antibody precipitated line in agar gel can be stained with Sudan black B; (7) thermostability: The antigenic properties in the serum are maintained at a temperature of 56° C. for 30 minutes; and (8) Form (as an antigen/antibody complex): the spherical particles are about 300 A in diameter.