Production of recombinant phage antibodies specific to gentamicin and their use in dot immunoassay

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Abstract

The potential of using phage display technology to obtain antigentamycin antibodies has been demonstrated. Antigentamycin recombinant antibodies were obtained for the first time using a sheep phage library (Griffin.1, UK). The interaction between obtained phage antibodies and gentamicin was monitored using a circular dichroism spectroscopy. It was shown that the interaction between antigentamycin phage antibodies and corresponding antibiotic is characterized by the presence of a characteristic exciton doublet: a positive peak at ~233 nm and a more intense negative peak with a maximum at ~240 nm. The possibility of gentamicin indication using a test system based on the dot immunoassay method and antigentamycin recombinant antibodies in aqueous solutions has been demonstrated for the first time; the lower limit of antibiotic detection is 0.5 μ g/ml. Using the dot immunoassay method, it was found that antigentamycin phage antibodies did not exhibit activity towards ampicillin and tetracycline, but showed activity towards kanamycin (lower limit of detection – 1 μ g/ml). The results are promising for further development of methods for gentamicin detection using recombinant phage antibodies.

About the authors

O. I. Guliy

Institute of Biochemistry and Physiology of Plants and Microorganisms – Subdivision Research Institution Saratov Federal Scientific Centre of the Russian Academy of Sciences

Email: guliy_olga@mail.ru
Saratov, 410049 Russia

D. S. Chumakov

Institute of Biochemistry and Physiology of Plants and Microorganisms – Subdivision Research Institution Saratov Federal Scientific Centre of the Russian Academy of Sciences

Email: guliy_olga@mail.ru
Saratov, 410049 Russia

V. S. Grinev

Institute of Biochemistry and Physiology of Plants and Microorganisms – Subdivision Research Institution Saratov Federal Scientific Centre of the Russian Academy of Sciences; Institute of Chemistry, N. G. Chernyshevsky Saratov National Research State University

Email: guliy_olga@mail.ru
Saratov, 410049 Russia; Saratov, 410012 Russia

O. A. Karavaeva

Institute of Biochemistry and Physiology of Plants and Microorganisms – Subdivision Research Institution Saratov Federal Scientific Centre of the Russian Academy of Sciences

Author for correspondence.
Email: guliy_olga@mail.ru
Saratov, 410049 Russia

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