Development and characterization of antibodies against aflatoxins

Abstract

A panel of ten monoclonal antibodies to 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 domains 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.

Development and properties of recombinant proteins based on the broadly neutralizing antibody to influenza A virus

“…Сборку бицистронной экспрессионной плазмиды для получения Fab-фрагмента антитела FI6 в клет-ках E. coli, биосинтез и очистку белка осуществляли по схеме, описанной ранее [6,7]. Полученные нук леотидные последовательности вариабельных доменов объединяли с константными доменами иммуноглобулина G человека (IgG1, каппа).…”

One-Step Detection of Aflatoxin-B1 Using scFv-Alkaline Phosphatase-Fusion Selected from Human Phage Display Antibody Library

“…Since obtaining specific antibody to free hapten is often very difficult with either naïve libraries or libraries derived from immunized animals, many strategies have been developed to increase the chance of selecting specific antibodies from a phage display antibody library. These include modification of the elution step by competing with free hapten [18], alternative hapten-carrier conjugates [6], selection from a focused library that favors binding haptens (cavity library) [19], or selection from an immunized library [20]. However, in this study, we were able to isolate specific antibody by using a simple bio-panning procedure.…”

Neutralizing monoclonal and chimeric antibodies to human interferon-γ

“…Purified mAbs were prepared from ascites fluid by affinity chromatography using protein A Sepharose as described previously [23].…”