Applying nanobody Phage Disolay technology to distinguish between brucella and yersinia lipopolysaccharides
2010
Naoufi, Alia | Khdrawi, A. | Mourad, A. | Abbady, A.
Brucellosis is a zoonotic disease caused by negative gram bacteria belonging to the genus Brucella. These bacteria contain in their outer cell wall a thick layer of lipopolysaccharides (LPS). LPS considered a very important elements for Brucella pathogenicity and immunogenicity. Furthermore, LPS appear a high similarity with Yersinia serotype 0:9 which can reach 98% in their internal structure, mainly the 0 chain component, causing regularly serum cross reactivity and confusion in results reading and analysis. Nanobody phage display technology is considered as a recent molecular biology technique performed by means of the genetic engineering of special type of antibodies, existing exclusively in Camelidea. It enables the obtaining of small binders, referred to as Nanobodies (Nb). Nbs, characterized with their high stability and solubility, are produced by gene expression in E. coli. In fact, they are extracted form a gene library of a high diversity, from which active binders are isolated by panning with phage display. In the early steps of this procedure, Nanobodies are present as displayed molecules on the surface of phages, or Nanobody-displaying phages (Nb-phages). In this work, a previously established Nanobody library, created from Arabian camel immunized with Brucella melitensis total antigens, was subjected to a subtractive panning with LPS from Yersinia followed with those from Brucella. This resulted in several Nb-phages able to distinguish efficiently between the LPS of these two genus. Results of this work could be invested in the development of precise and rapid diagnostic kits to discriminate between Brucella and Yersinia infections.
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