反应详情

To a stirred solution of 16 mg of sheep anti-digoxin in 10 ml of pH 8.0 phosphate buffered saline at 0° was added dropwise 0.1 ml of a 0.1M solution of gallocyanine NHS ester (prepared by a procedure similar to that of Example 4 for the preparation of the NHS ester of DECAS) in dry DMF over a five minute period. After stirring at 0°-4° for 18 hours, 0.1 ml of 4M hydroxylamine hydrochloride was added all at once and stirring was continued for thirty minutes at 0°-4°. The blue solution was then purified by gel filtration, dialysis, and centrifugation to give 7.6 ml of 1.44 mg/ml blue antibody solution. The solution contained an average of 2.6 moles gallocyanine per mole of anti-digoxin. The absorption maxima of the conjugate were at 278 nm and 625 nm, and the relative quantum yield of the conjugate was determined to be 0.02.

1. customprepared by a procedure similar to that of Example 4
2. stirringstirring
3. waitwas continued for thirty minutes at 0°-4°
4. filtrationThe blue solution was then purified by gel filtration, dialysis, and centrifugation
5. customto give 7.6 ml of 1.44 mg/ml blue antibody solution