ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)
"Biotechnologia Acta" v. 6, no. 4, 2013
Р. 19-32, Bibliography 16, Russian
Universal Decimal classification: 602.68:615.372
OBTAINING OF MONOCLONAL ANTIBODIES AGAINST CHOLERA TOXIN AND HEAT LABILE ENTEROTOXIN OF E. coli FOR DEVELOPMENT OF THE TOXINS DIPLEX ANALYSIS IN ENVIRONMENTAL SPECIMENS
Eu. V. Grishin,T. I. Valiakina
Federal State Institution of Science
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry
of Russian Academy of Sciences,Moscow, Russian Federation
The present study focuses on development of monoclonal antibodies (MAbs) which specifically interact with cholera toxin or heat labile enterotoxin of E. coli. Such monoclonal antibodies MAbs are possessed of ability to identify cholera toxin or heat labile enterotoxin in different immunochemical assays. We obtained hybridoma clones which produced monoclonal antibodies of IgG isotypes to cholera toxin and heat labile enterotoxin. On application of the method of serial dilutions we selected the clones which produced monoclonal antibodies with specific activity against only one of the toxins. We found the 16 pairs of monoclonal antibodies to cholera toxin and 28 ones to heat labile enterotoxin. By means of these monoclonal antibodies it was possible to realize the quantitative analysis of theses toxins in sandwich immunoassay ELISA and diplex sandwich xMAP-assay. The limits of detection of cholera toxin and heat labile enterotoxin in ELISA in control buffer were 0.2 and 0.4 ng/ml, respectively, and in xMAP assay — 0.01 and 0.08 ng/ml, respectively. In probes of cow milk, meat soup, pond water and nasopharyngeal washes cholera toxin was detected in the both assays with the same limits of detections, but heat labile enterotoxin limits of detections were above the ones in control buffers.
Key words: ELISA, sandwich assay, multiplex immunoassay, xMAP analysis, monoclonal antibodies, minimal detectable concentration, cholera toxin, heat labile enterotoxin of E. coli.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2013
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