Conjugation of enzymes to immunoglobulins using dimaleimides.
A method is described for coupling enzymes to immunoglobulins using sulphydryl derivatives of the proteins and a dimaleimide which is relatively water-soluble. Parameters affecting the performance of the conjugates have been examined including level of sulphydryl incorporation, ratio of enzyme/immunoglobulin and nature of dimaleimide used. Peroxidase-immunoglobulin conjugates made by the dimaleimide method have been compared with those made by the periodate oxidation method and found to have a superior performance. Immunoglobulin has been linked to peroxidase (horseradish peroxidase, EC 1.11.1.7), glucose oxidase from Aspergillus niger, (EC 1.1.3.4), penicillinase from Bacillus cereus beta-lactamase I (EC 3.5.2.6), and beta-galactosidase from Escherichia coli (EC 3.2.1.23).
D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G; D007136:Immunoglobulins; D008301:Maleimides; D010544:Peroxidases
D007074:Immunoglobulin G; D007136:Immunoglobulins; D008301:Maleimides; C011408:bis(N-maleimidomethyl)ether; C011328:N,N'-4-phenylenedimaleimide; D010544:Peroxidases