In silico peptide prediction for antibody generation to recognize 5‐enolpyruvylshikimate‐3‐phosphate synthase (EPSPS) in genetically modified organisms

Abstract

For the prospective immunorecognition of 5-enolpyruvylshikimate-3-phosphate synthase (CP4-EPSPS) as a biomarker protein expressed by transgenic soybean, an extensive in silico evaluation of the referred protein was performed. The main objective of this study was the selection of a set of peptides that could function as potential immunogens for the production of novel antibodies against CP4-EPSPS protein. For this purpose, the protein was in silico cleaved with trypsin/chymotrypsin and the resultant peptides were extensively analyzed for further selection of the best candidates for antibody production. The analysis enabled the successful proposal of four peptides with potential immunogenicity for their future use as screening biomarkers of genetically modified organisms. To our knowledge, this is the first attempt to select and define potential linear epitopes for the immunization of animals and, subsequently, to generate adequate antibodies for CP4-EPSPS recognition. The present work will be followed by the synthesis of the candidate peptides to be incubated in animals for antibody generation and potential applicability for the development of an immunosensor for CP4-EPSPS detection.

Cry1A(b)16 toxin from Bacillus thuringiensis: Theoretical refinement of three‐dimensional structure and prediction of peptides as molecular markers for detection of genetically modified organisms

“…), on the basis of their sequence and immunogenicity . The parameters used in silico involved theoretical digestion at trypsin and chymotrypsin cleavage sites, sequence and structure determination of epitopes (B‐Cell and T‐cell epitope prediction tools), cross‐reactivity, and secondary‐structure prediction for the selected peptides …”

Polyclonal antibody production anti Pc_312-324 peptide. Its potential use in electrochemical immunosensors for transgenic soybean detection

“…Pc_312-324 Peptide (PEP), whose amino acid sequence is shown in Schematic 1, was selected by in silico evaluation based on their sequence and immunogenicity [28] Peptide was manually synthesized using a solid phase approach with Fmoc/t-butyl chemistry [29]. Peptide elongation was carried out in polypropylene syringes fitted with a polyethylene porous disk.…”

“…al. proposed to synthesize a set of peptides that could be potential antigens for the production of new antibodies against CP4-EPSPS protein present in transgenic soybean by using bioinformatics [28].…”

“…In this work, we produced a new polyclonal antibody by using one of these candidates peptides proposed by Marani et al [28] synthesized in our laboratory, to immunize the animals. This antibody was then probed to recognize the CP4-EPSPS protein, while linked into an electrode surface to evaluate the potential use in an electrochemical immunosensor for CP4-EPSPS sensitive, rapid and specific detection.…”

Mass spectrometry combined with affinity probes for the identification of CP4 EPSPS in genetically modified soybeans

“…[2][3][4] Soybean continues to be the dominant GM crop, occupying almost 49% (85 million hectares) of the world's total GM planted areas (more than 175 million hectares). 3,5 However, consumer concerns about the safety of GMOs have prompted the development of GMO food labeling legislation. Labeling thresholds have been established by several countries to strengthen and standardize the development of GMOs.…”