Process for coupling biological substances by covalent bonds
First Claim
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1. In a process for coupling at least two biological substances by covalent bonds by means of a cross-linking agent, the said cross-linking agent being benzoquinone, the improvement comprising the steps of:
- (a) treatment of Fab fragment of an immunoglobulin as the first substance dissolved in solution by at least about 200 times its molar amount of benzoquinone, said reaction known as the activation reaction being performed in a homogeneous liquid medium;
(b) removal of the reaction products of step (a) and the excess benzoquinone and recovery of the activated Fab fragment;
(c) contacting the activated Fab fragment with the second substance to be coupled.
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Abstract
A process for coupling biological substances by covalent bonds that involves the use of benzoquinone as cross-linking agent in large excess compared with the substance to be activated. The activation reaction is realized in a homogeneous liquid medium. The process makes it possible, for example, to couple antibodies to enzymes for determining or detecting said antibodies.
The process can be used for the determination of antitetanic antibodies in human serum.
115 Citations
13 Claims
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1. In a process for coupling at least two biological substances by covalent bonds by means of a cross-linking agent, the said cross-linking agent being benzoquinone, the improvement comprising the steps of:
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(a) treatment of Fab fragment of an immunoglobulin as the first substance dissolved in solution by at least about 200 times its molar amount of benzoquinone, said reaction known as the activation reaction being performed in a homogeneous liquid medium; (b) removal of the reaction products of step (a) and the excess benzoquinone and recovery of the activated Fab fragment; (c) contacting the activated Fab fragment with the second substance to be coupled. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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