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Information × Registration Number 0211U013200, 0110U006124 , R & D reports Title Developing a fundamental basis for obtaining new generation immunoreagents popup.stage_title Head Pavlova Maryna Valeriivna, Registration Date 10-01-2012 Organization Institute of Molecular Biology and Genetics popup.description2 We aimed at construction of the chimeric protein scFv-BАРmut comprises single-chain antibody (single-chain fragment variable, scFv) and engineered Escherichia coli alkaline phosphatase moieties (bacterial alkaline phosphatase mutated, BАРmut) and /or fluorescent proteins, as well, as its obtaining by bacterial synthesis and characteristics. Engineered scFv-BАРmut and ScFv-mCherry proteins were produced in E. coli. Amino acid substitutions D330N and D153G resulted in a significant increase of the catalytic activity of the engineered phosphatase moiety compared to the wild-type protein. We demonstrate high specificity of the interaction between scFv-BАРmut and its cognate antigen and potential applicability of crude bacterial periplasm and culture medium fractions for direct one-stage ELISA and western blotting-based immunoaqssays. The scheme of ScFv-mCherry refolding was created. The convenience of designing the sequence encoding specific scFv, simplicity of obtaining recombinant protein by bacterial synthesis, high sensitivity to antigen, as well as the wide spectrum of the available colorimetric, luminescent, and fluorescent substrates for the phosphatase, as well as brightness of mCherry, promise new potential perspectives and utility of the obtained chimeric proteins. Product Description popup.authors Okunev O. V. Pavlova M. V. Shpileva S. P. popup.nrat_date 2020-04-02 Close