DEVELOPMENT OF A MULTIEPITOPE RECOMBINANT PROTEIN PRODUCTION STRATEGY FOR VIRAL DIAGNOSIS

ROTA PR; LORCH MS; COLLADO MS; GOÑI SE

Mar del Plata

Congreso; LI Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2015

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

The goal of this project is the production of recombinant antigens to be used in the serological diagnosis of different Flavivirus and Alphavirus, with the overall aim of improving its diagnosis and epidemiological surveillance. Currently, the diagnostic of these viruses rely on the use of whole virus antigens and are consequently associated with false positives due to serologic cross-reactivity, high cost of antigen production, and biohazard risk. This has prompted the need to develop an alternate antigen to replace the whole virus in diagnostic. We have designed and expressed a novel recombinant protein antigen by assembling linear IgG-specific SLEV or VEEV epitopes, chosen on the basis of computer predictions and bibliographic data of related viral species. The recombinant virus multiepitope protein was expressed in Escherichia coli and fused to a his-tag tail as a purification method. Also it will be expressed as a fusion protein using GST, maltose or tioredoxin. Finally we planed develop an in-house ELISA to detect anti-SLEV or anti-VEEV antibodies in a panel of positive and negative sera using the purified recombinant multiepitope protein as the capture antigen. The selection of epitopes and the use of E. coli as expression system, along with simple purification, have the potential to lead to the development of an inexpensive, highly specific and sensitive diagnostic test.