Dimethyl Suberimidate as an Effective Crosslinker for Antibody-Enzyme Conjugation
- 1 December 1991
- journal article
- research article
- Published by Taylor & Francis in Preparative Biochemistry
- Vol. 21 (4) , 215-227
- https://doi.org/10.1080/10826069108018574
Abstract
Dimethyl suberimidate (DMS), a bifunctional reagent was used for the first time to crosslink the α -feto protein monoclonal antibodies (AFPMAb) to horse raddish peroxidase (HRP). Three batches of conjugates were prepared, purified by Sephadex gel chromatography and evaluated for their immunological reactivity. The Rz values obtained for AFPMAb-HRP conjugate were 0.39 to 1.36. Under optimised conditions the ELISA results showed the optical density of 1.9. The iso-electric focusing for the conjugate revealed different degrees of crosslinking between antibodies and HRP. It was evident that isoperoxidase-C was involved in the crosslinking process. From the dot ELISA as low as 25 pg of AFP in the test samples could be detected with AFPMab-HRP conjugate. The conjugate prepared by DMS was stable at 0°C for more than 10 months.Keywords
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