Isolation of different types of manobody displying phages capable of distinguishing between the most two important species of brucella, B. melitensis and B. abortus, in Syria
2010
Khdrawi, A. | El-Naoufi, A. | Al-Mariri, A.Q. | Abbady, A.
Nanobody phage display technology is considered as recent molecular biology technique performed through the genetic engineering of special type of antibodies existing exclusively in Camelidea. It enables the obtaining of small binders, referred to as Nanobodies, conferred with high stability and solubility and was produced by gene expression system in E. coli. Nanobodies are extracted form gene library of 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). This work was established on Nanobody library previously created from Arabian camel immunized with Brucella melitensis total antigens. It is a member of the genus Brucella which is considered as a very important infectious agent because of its impact on human and animal health with the concomitant economical loses in Syria. Locally, cases of infection with B. melitensis are more redundant than with B. abortus. Distinguishing between these two types of bacteria is still considered as a problem because of the big similarity in their outer structure and protein content. In this work, a subtractive panning with B. abortus antigens followed with those from B. melitensis was performed on the Nanobody library resulting in several Nbphages able to distinguish efficiently between these two species of Brucella. Results of this work could be invested in the development' of precise and rapid diagnostic kits for detecting Brucellosis and defining its responsible type of bacteria
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