期刊
ACTA NATURAE
卷 3, 期 3, 页码 85-92出版社
RUSSIAN FEDERATION AGENCY SCIENCE & INNOVATION
DOI: 10.32607/20758251-2011-3-3-85-92
关键词
horseradish peroxidase; antibodies; recombinant conjugates; Pichia pastoris expression
资金
- German Federal Ministry of Science and Technology (BMBF) [0311574]
Recombinant immunoconjugates of marker enzymes with antigens or antibodies present considerably more advantages than those obtained by conventional methods of chemical synthesis; i.e. they are homogeneous, have a strictly determined stoichiometry, and retain the functional activity of both a marker protein and an antigen/antibody. Based on the pPICZ alpha B shuttle vector, we first managed to obtain a recombinant conjugate of key marker enzyme horseradish peroxidase (HRP) with Fab fragments of antibodies against atrazine. The resulting genetic construction allows us to switch to any other antibody sequence, via the simple re-cloning of variable parts and an additional reporter enzyme. Conjugates were successfully produced in the Pichia pastoris methylotrophic yeast expression system. The target activity of the conjugates (both enzymatic and antigen-binding) has been demonstrated by ELISA method.
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