首页|CD36蛋白B细胞抗原表位的生信预测及其多抗原肽的制备

CD36蛋白B细胞抗原表位的生信预测及其多抗原肽的制备

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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。
Prediction of B cell epitopes of CD36 and preparation of MAP
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.

plateletCD36B cell epitopeMAP

刘静、徐秀章、丁浩强、邓晶、王嘉励、陈扬凯、夏文杰、叶欣

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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

血小板 CD36 B细胞表位 MAP肽

广州市科技计划项目广州市科技计划项目广州市科技计划项目广州市科技计划项目广东省基础与应用基础研究基金自然科学基金项目

2023A03J09972024A03J02272023A03J05522023A03J5492024A1515012652

2024

中国输血杂志
中国输血协会 中国医学科学院输血研究所

中国输血杂志

CSTPCD
影响因子:1.279
ISSN:1004-549X
年,卷(期):2024.37(8)