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Development and characterization of antibodies against aflatoxins

Authors
  • Kalinichenko, A. A.1, 2
  • Toporova, V. A.1
  • Panina, A. A.1
  • Aliev, T. K.1, 3
  • Kryukova, E. A.1
  • Shemchukova, O. B.4
  • Solopova, O. N.4
  • Pozdnyakova, L. P.4
  • Sveshnikov, P. G.4
  • Dolgikh, D. A.1, 2
  • Kirpichnikov, M. P.1, 2
  • 1 Russian Academy of Sciences, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, ul. Miklukho-Maklaya 16/10, Moscow, 117997, Russia , Moscow (Russia)
  • 2 Moscow State University, Faculty of Biology, Moscow, 119899, Russia , Moscow (Russia)
  • 3 Moscow State University, Faculty of Chemistry, Moscow, 119899, Russia , Moscow (Russia)
  • 4 Russian Research Center for Molecular Diagnostics and Therapy, Moscow, Russia , Moscow (Russia)
Type
Published Article
Journal
Russian Journal of Bioorganic Chemistry
Publisher
SP MAIK Nauka/Interperiodica
Publication Date
Jan 01, 2010
Volume
36
Issue
1
Pages
114–123
Identifiers
DOI: 10.1134/S1068162010010139
Source
Springer Nature
Keywords
License
Yellow

Abstract

A panel of ten monoclonal antibodies against aflatoxins B1, B2, and G2 was produced and comprehensively characterized. The affinity and cross reactivity of these antibodies were determined using the methods of direct, indirect, and competitive ELISA. The structures of monoclonal antibody genes were comprehensively studied and the variable and constant regions of the antibody genes were cloned and sequenced. Sequencing analysis confirmed the results of isotyping the light and heavy antibody chains obtained by ELISA. Variable and constant fragments of the antibody genes were cloned into a bicistron expression vector for the recombinant Fab-fragment for one of the antibodies expressed in Escherichia coli and purified. Thus, data were obtained that can be useful for the development of an aflatoxin detection system on the basis of the described monoclonal antibodies and the creation of recombinant antibodies with changed parameters of specificity using protein engineering methods.

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