CD36蛋白B细胞抗原表位的生信预测及其多抗原肽的制备  

Prediction of B cell epitopes of CD36 and preparation of MAP

作　　者：刘静[1,2,3] 徐秀章[1,2,3] 丁浩强 邓晶[1,2,3] 王嘉励 陈扬凯[1,2,3] 夏文杰 叶欣[1,2,3] LIU Jing;XU Xiuzhang;DING Haoqiang;DENG Jing;WANG Jiali;CHEN Yangkai;XIA Wenjie;YE Xin(Guangzhou Blood Center,Guangzhou 510091,China;Institute of Blood Transfusion and Hematology at Guangzhou Medical University;Guangzhou Key Laboratory of Blood Safety)

机构地区：[1]广州血液中心,广东广州510091 [2]广州医科大学输血与血液病研究所 [3]广州市血液安全重点实验室

出　　处：《中国输血杂志》2024年第8期853-858,共6页Chinese Journal of Blood Transfusion

基　　金：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)。

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 antibodies using MAP with B cell epitopes.Methods The potential B cell epitopes of CD36 were analyzed by bioinformatics methods,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 prepared.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 containing CD36 B cell epitopes can provide the experimental basis for the preparation and related research of CD36 antibodies.

关 键 词：血小板 CD36 B细胞表位 MAP肽 

分 类 号：R457[医药卫生—治疗学]