目的:采用生物信息学技术,对前期研究获得的卵泡刺激素受体(FSHR)的B细胞表位肽段进一步筛选、优化,以获得理想的FSHR靶标抗原。方法:①利用Insight II分子模拟软件寻找FSHR与FSH相互作用的区域。②应用DNA STAR软件中的Protein模块对优化后抗原进行综合分析。③肽结合实验对优化后抗原进行免疫原性检测。结果:≥5个氨基酸片段的柔性区域位于FSHR第30-44、52-65、112-121、174-182、190-210、223-245区段。当FSHR胞外区第37位缬氨酸被赖氨酸或谷氨酸替代时,具有较好的亲水性及抗原性。而肽结合实验证实肽32-44具有更好的免疫原性。结论:FSHR胞外区肽段32-44有可能成为避孕疫苗的理想抗原。
Objective: To obtain the desired antigen targeting - FSHR, we screened and optimized the B cell epitope for the extra- cellular domain of the follicle stimulating hormone receptor (FSHR) . Methods: (1)Insight II molecular simulation software was used to identity amino acid sites of interaction interface between FSH and FSHR. (2)The secondary structure and surface properties of the extracellular domain of FSHR, such as physical and chemical properties, hydrophilicity, antigenicity and plasticity, were analyzed by a variety of meth- ods. (~)The peptide of the epitopes was synthesized and used for coating streptavidin plate. The immunogenicity of the peptides was deter- mined. Results: Many distinct antigenic epitopes in the extracellular domain of FSHR were identified by computation, the possible location of which was in the regions of 30 - 44, 52 - 65, 112 - 121, 174 - 182, 190 - 210 and 223 - 245. When the FSHR ectodomain 37th valine was replaced by lysine and glutamic acid, the antigen had better hydrophilicity and antigenic. However, the peptide 32 - 44 was confirmed the best immunogenicity in ELISA assay. Conclusion: FSHR ectodomain peptide 32 -44 can be the ideal contraceptive vaccine antigens.