Tresyl-activated support for high-performance affinity chromatography.
A new activated support TSK gel Tresyl-5PW was evaluated for the coupling of antibodies, which was found to occur easily under mild conditions with high yields. Optimum coupling conditions were a 2-h reaction at 25 degrees C in 1 M phosphate buffer (pH 7.5) when 2-3 mg antibody/ml support is to be coupled and a 6-7-h reaction when ca. 10 mg antibody/ml support is to be coupled. When antibodies were coupled under these conditions, antibody coupling yields greater than 80% and antigen binding efficiencies of 70-80% were achieved, probably owing to a selective attachment of the Fc region of the antibodies. Antigens (human serum proteins) could be separated rapidly without denaturation on antibody-coupled Tresyl-5PW.
Central Research Laboratory, TOSOH Corporation, Yamaguchi, Japan.
K
K
Nakamura
T
T
Hashimoto
Y
Y
Kato
K
K
Shimura
K
K
Kasai
D000906:Antibodies / Q000302:isolation & purification; D001798:Blood Proteins / Q000276:immunology / Q000302:isolation & purification; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D005782:Gels; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000302:isolation & purification; D008970:Molecular Weight; D013450:Sulfones
D000906:Antibodies; D001798:Blood Proteins; D005782:Gels; D007074:Immunoglobulin G; D013450:Sulfones; C032605:2,2,2-trifluoroethanesulfonyl chloride