RhD多肽抑制抗原抗体结合的效果*

doi:10.3969/j.issn.1671-2587.2020.01.006

临床输血与检验 ›› 2020, Vol. 22 ›› Issue (1): 23-27.DOI: 10.3969/j.issn.1671-2587.2020.01.006

RhD多肽抑制抗原抗体结合的效果*

吴凡, 庄乃保, 梁爽, 梁延连, 苏宇清

518035 深圳市血液中心

收稿日期:2018-12-07 出版日期:2020-02-20 发布日期:2020-02-28
作者简介:吴凡(1982-),女,广东梅州人,副主任技师,硕士,主要从事输血技术学研究工作,(Email)fancy_32fan@126.com。
基金资助:
*本研究受深圳市卫生计生系统项目公共卫生防控策略研究项目(No.SZGW2018008);深圳市基础研究自由探索项目(No.JCYJ20160427172335974);深圳市基础研究学科布局项目(No.JCYJ20170412150719814)资助

The Effect of RhD Polypeptide to Inhibit the Reaction of RhD Antigen and Anti-D

WU Fan, ZHUANG Nai-bao, LIANG Shuang, et al

Shenzhen Blood Center,Shenzhen 518035

Received:2018-12-07 Online:2020-02-20 Published:2020-02-28

摘要/Abstract

摘要： 目的评价RhD多肽抑制抗-D抗体与RhD抗原结合的效果。方法根据随机噬菌体展示文库筛选得到RhD抗原模拟多肽的氨基酸序列,合成RhD多肽。利用生物信息学技术分析多肽理化性质及二级结构,微柱凝胶低离子抗人球蛋白卡凝集抑制试验验证有效多肽,显微镜观察多肽有效抑制浓度,流式细胞仪法检测多肽抑制效果。结果 Peptide 4噬菌体多肽最为稳定,未形成无规则卷曲与抗原性,且抑制IgG型单克隆抗-D与RhD抗原的反应效果较为明显,显微镜观察呈一定的剂量效应。流式细胞仪法显示当Peptide 4浓度达2.5 mg/50μL时对IgG型单克隆抗-D抗体与O型RhD阳性红细胞的凝集具有抑制作用。结论 Peptide 4为有效多肽,对抑制抗-D与RhD抗原的凝集具有一定效果,对RhD蛋白结构与功能研究及抗原性改造,提高临床输血的安全性等具有重要意义。

关键词: Rh血型, 多肽, 抗-D, D抗原

Abstract: Objective To evaluate the RhD polypeptide's inhibitory effect of the reaction of RhD antigen and anti-D. Methods RhD phage polypeptides were synthesized according to the amino acid sequence from phage display library screening. The physical and chemical properties and secondary structure of polypeptides were analyzed by bioinformatics. The effective polypeptides were verified by agglutination inhibition experiment using microcolumn gel (LISS and anti-human globulin). The effective blocking concentration of polypeptides was observed by using microscope to observe the agglutination reaction. The blocking effect of polypeptides was detected by flow cytometry. Results Peptide 4 is the most stable polypeptide. There are no random coil and antigenic in this polypeptide. Peptide 4 is able to inhibit the agglutination of IgG monoclonal anti-D and D+ cells in a concentration dependent manner and completely inhibit the reaction when the concentration reached 2.5 mg/50 μL. Conclusion Peptide 4 is the effective polypeptide. It is able to block the binding sites of anti-D and RhD antigen. The result has great significance to study the structure and function of RhD protein, to modify the antigen of RhD protein, and to improve the safety of clinical blood transfusion.

Key words: Rh blood group, Polypeptide, Anti-D antibody, D antigen

中图分类号:

R446.63

吴凡, 庄乃保, 梁爽, 梁延连, 苏宇清. RhD多肽抑制抗原抗体结合的效果*[J]. 临床输血与检验, 2020, 22(1): 23-27.

WU Fan, ZHUANG Nai-bao, LIANG Shuang, et al. The Effect of RhD Polypeptide to Inhibit the Reaction of RhD Antigen and Anti-D[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2020, 22(1): 23-27.

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RhD多肽抑制抗原抗体结合的效果*

The Effect of RhD Polypeptide to Inhibit the Reaction of RhD Antigen and Anti-D

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