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CD36蛋白B细胞抗原表位的生信预测及其多抗原肽的制备OACSTPCD

Prediction of B cell epitopes of CD36 and preparation of MAP

中文摘要英文摘要

目的 分析CD36 蛋白的结构,寻找可能的B细胞抗原表位,制备含有B细胞表位的多抗原肽(multiple antigenic peptide,MAP),为基于B细胞表位的MAP用于CD36 抗体的制备提供前期实验基础.方法 通过生物信息学方法分析CD36 蛋白的理化性质、二级结构、潜在磷酸化及糖基化位点等,预测可能的B细胞表位.以多聚赖氨酸为核心基质,采用Fmoc方法合成含有CD36 B细胞表位的八分枝MAP,并利用反向高效液相色谱(RP-HPLC)分析MAP的纯度、质谱分析法测定MAP的分子量,对合成的CD36-MAP进行鉴定.结果 CD36 是 1 种稳定的、亲水性的碱性蛋白,二级结构以无规则卷曲为主,具有多个磷酸化、糖基化位点,抗原性较强.综合分析获得可能的优势B细胞表位 4 个,制备了 4 个含有优势B细胞表位的MAP,RP-HPLC分析表明合成的MAP 的纯度均在 85%以上,其中 3 个MAP的分子量与理论预期值相符.结论 CD36 具有较强的抗原性,利用预测得到的 4 个可能的B细胞表位合成MAP,为CD36 抗体的制备和相关研究提供实验基础.

Objective To analyze the structure of CD36,search the possible B cell epitopes and prepare multi-antigen peptides(MAP)with B cell epitopes,so as to provide a preliminary experimental basis for the preparation of CD36 antibod-ies using MAP with B cell epitopes.Methods The potential B cell epitopes of CD36 were analyzed by bioinformatics meth-ods,including physical and chemical properties,secondary structure,potential phosphorylation and glycosylation sites.Eight-branch MAP with CD36 B cell epitopes were synthesized by FMOC using polylysine as the core matrix.The purity of MAPs was analyzed by reverse high-performance liquid chromatography chromatography(RP-HPLC),and the molecular weight of MAPs was determined by mass spectrometry.Results CD36 is a stable and hydrophilic alkaline protein,with multiple phosphorylation and glycosylation sites and strong antigenicity,and its secondary structure is mainly characterized by irregular curls.Four potential B cell epitopes were obtained and 4 MAPs containing potential B cell epitopes were pre-pared.RP-HPLC analysis showed that the purity of the MAPs were above 85%,and the molecular weight of 3 MAPs was consistent with the expected theoretical molecular weight.Conclusion CD36 on platelet has strong antigenicity.MAPs con-taining CD36 B cell epitopes can provide the experimental basis for the preparation and related research of CD36 antibodies.

刘静;徐秀章;丁浩强;邓晶;王嘉励;陈扬凯;夏文杰;叶欣

广州血液中心,广东 广州 510091||广州医科大学输血与血液病研究所||广州市血液安全重点实验室

临床医学

血小板CD36B细胞表位MAP肽

plateletCD36B cell epitopeMAP

《中国输血杂志》 2024 (008)

853-858 / 6

广州市科技计划项目(2023A03J0997、2024A03J0227,2023A03J0552,2023A03J549);广东省基础与应用基础研究基金自然科学基金项目(2024A1515012652)

10.13303/j.cjbt.issn.1004-549x.2024.08.002

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