Generation and characterization of biotinylated recombinant Fab antibody fragment against cortisol

This paper describes the in vivo generation method of biotinylated recombinant Fab antibody fragments. The original molecular vectors for Escherichia coli Fab fragments expression were designed. In vivo biotinylated recombinant antibody Fab fragment against cortisol was generated. The kinetic parame...

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Bibliographic Details
Published in:Russian journal of bioorganic chemistry 2016, Vol.42 (1), p.22-28
Main Authors: Dormeshkin, D. O., Kuprienko, O. S., Svirid, A. V., Gilep, A. A., Sviridov, O. V., Usanov, S. A.
Format: Article
Language:English
Subjects:
Biochemistry
Biomedical and Life Sciences
Biomedicine
Bioorganic Chemistry
Escherichia coli
Life Sciences
Organic Chemistry
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Summary:This paper describes the in vivo generation method of biotinylated recombinant Fab antibody fragments. The original molecular vectors for Escherichia coli Fab fragments expression were designed. In vivo biotinylated recombinant antibody Fab fragment against cortisol was generated. The kinetic parameters of interaction of these antibody fragments with cortisol-BSA complex were measured via the biolayer interferometry method. An equilibrium dissociation constant of this interaction K D is 5.48 × 10 –10 M. The interaction is reversible and it could be competitively inhibited by free cortisol addition. These results could be used in generating of immunoassays for quantitative cortisol determinations. The in vivo biotinylation system under review is universal and suitable for expression of any biotinylated Fab fragments in the E. coli system.
ISSN:1068-1620
1608-330X
DOI:10.1134/S1068162016010039