Marani Mariela M, Costa Joana, Mafra Isabel, Oliveira Maria Beatriz P P, Camperi Silvia A, Leite José Roberto de Souza Almeida
CENPAT-CONICET, Centro Nacional Patagónico, Consejo Nacional de Investigaciones Científicas y Técnicas, Puerto Madryn, Argentina.
Biopolymers. 2015 Mar;104(2):91-100. doi: 10.1002/bip.22609.
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.
为了前瞻性地免疫识别5-烯醇丙酮酰莽草酸-3-磷酸合酶（CP4-EPSPS）作为转基因大豆表达的生物标志物蛋白，对该蛋白进行了广泛的计算机模拟评估。本研究的主要目的是选择一组肽，这些肽可作为潜在免疫原，用于生产针对CP4-EPSPS蛋白的新型抗体。为此，用胰蛋白酶/胰凝乳蛋白酶对该蛋白进行计算机模拟切割，并对所得肽进行广泛分析，以进一步选择用于抗体生产的最佳候选肽。该分析成功提出了四种具有潜在免疫原性的肽，它们未来可作为转基因生物的筛选生物标志物。据我们所知，这是首次尝试选择和定义用于动物免疫的潜在线性表位，并随后产生足够的抗体用于CP4-EPSPS识别。后续工作将合成候选肽，在动物体内孵育以产生抗体，并探讨其在开发用于CP4-EPSPS检测的免疫传感器方面的潜在适用性。
