Exploiting bioinformatic tools to assess CP4-EPSPS peptides as potential biomarkers of genetically modified soybean

COSTA JOANA B.; MARANI MARIELA M.; MAFRA ISABEL; OLIVEIRA MARÍA B.P.P.; CAMPERI SILVIA A.; LEITE JOSÉ R. S. A.

Porto

Conferencia; 5th MoniQA International Conference; 2015

MoniQA Association

The theoretical prediction of epitopes is a highly challenging task, although significant progress has been made in finding synthetic peptides that could mimic epitopes with potential application for diagnostic and research purposes. Presently, a large set of computational tools has become available for the prediction of linear and conformational epitopes in proteins, mostly based on T- and B-cell prediction softwares. So far, the use of bioinformatics to predict the potential immunogenicity of peptides has been mainly focused on medical purposes, with some successful applications already described in the development of vaccines. Using bioinformatics, the 5-enolpyruvulshikimate-3-phosphate synthase (CP4-EPSPS) protein, which is expressed by genetically modified soybean, was extensively in silico analysed. The resultant peptides were evaluated for further selection of the best candidate peptides to generate immunological response and subsequent production of anti-CP4-EPSPS antibodies. For this purpose, several available computational tools, namely DiscoTope [1], ElliPro [2] or POPI 2.0 [3] softwares, were used to evaluate a total of 32 sequences ranging from 10 to 25 amino acids in length, resulting from the in silico cleavage of CP4-EPSPS. The analysis enabled the successful proposal of four peptides, namely Pc_19-32, SGTVRIPGDKSISH; Pc_67-77, GARIRKEGDTW; Pc_141-156, GVQVKSEDGDRLPVTL and Pc_312-324, KGVTVPEDRAPSM for the prospective immunorecognition of CP4-EPSPS, thus facilitating the development of immunoassays for the detection of genetically modified soybean [4]. Additionally, considering that the expression of this protein is common to many other transgenic crops, these candidate peptides can be very useful as screening markers for the presence of GMO. In spite of the wide application of bioinformatics to medical areas, this is the first attempt to select and define potential linear epitopes for the immunisation of animals and subsequent production of anti-CP4-EPSPS antibodies. Actually, the selected peptides and respective antibodies are currently being synthesised aiming at developing an immunosensor for CP4-EPSPS detection.