人神经丝中分子量亚单位多肽的免疫原性及其抗体的特异性

doi:10.6040/j.issn.1671-7554.0.2017.843

山东大学学报 (医学版) ›› 2018, Vol. 56 ›› Issue (5): 58-63.doi: 10.6040/j.issn.1671-7554.0.2017.843

• • 上一篇

人神经丝中分子量亚单位多肽的免疫原性及其抗体的特异性

李昕¹,刘建伟¹,薛皓²,郑文慧¹,郑雯¹,秦泽明¹,温红玲¹,周晓莹³,王志玉¹,赵丽¹

1.山东大学公共卫生学院卫生微生物检验学系, 山东济南 250012;2.山东大学齐鲁医院神经外科, 山东济南 250012;3.山东大学齐鲁医院超声科, 山东济南 250012

收稿日期:2017-09-05 发布日期:2022-09-27
通讯作者: 赵丽. E-mail:dlzhl@sdu.edu.cn
基金资助:
山东省科技发展计划(2014GSF121014)

Human neurofilament medium polypeptide immunogenicity and specificity of antibodies

LI Xin¹, LIU Jianwei¹, XUE Hao², ZHENG Wenhui¹, ZHENG Wen¹, QIN Zeming¹, WEN Hongling¹, ZHOU Xiaoying³, WANG Zhiyu¹, ZHAO Li¹

1. Department of Laboratory Science of Sanitary Microbiology, School of Public Health, Shandong University, Jinan 250012, Shandong, China;
2. Department of Neurosurgery, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China;
3. Department of Ultrasound, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China

Received:2017-09-05 Published:2022-09-27

摘要/Abstract

摘要： 目的研究人神经丝中分子量亚单位多肽(NFM)的免疫原性及其抗体的特异性,为NFM单克隆抗体(McAb)的制备及ELISA检测方法的建立奠定基础。方法分析人、大鼠、小鼠NFM氨基酸序列,选择人NFM 4段肽P1、P2、P3、P4,人工合成并与匙孔槭血蓝蛋白(KLH)偶联,免疫小鼠收获血清,ELISA检测抗体效价,用Western blotting及免疫组化(IHC)检测各抗体与人、大鼠、小鼠及体外表达的NFM的反应性。结果 4种多肽具有较强的免疫原性,产生了高效价抗体,IHC结果显示,4种抗体均能与人脑组织胶质瘤、大鼠和小鼠脑组织中的NFM反应,Wesntern blotting结果显示,抗P1、P4抗体可与人脑组织胶质瘤、大鼠和小鼠脑组织中NFM反应,抗P1抗体出现2个条带;抗P1、P2、P3抗体均能与原核表达的NFM反应。结论所选择4段多肽具有较好的免疫原性,小鼠所产生抗体能与人、大鼠和小鼠脑组织及体外表达的NFM反应,可进一步用于NFM McAb的制备。

关键词: 中分子量神经丝亚单位, 多肽, 免疫原性, 抗体特异性

Abstract: Objective To explore the immunogenicity of human middle subunit of neurofilament(NFM)polypeptide and the specificity of the antibodies in order to provide basis for the preparation of monoclonal antibody(McAb)and establishment of an ELISA detection method. Methods The amino acid sequences of NFM in humans, rats and mice were analyzed. Four polypeptides(P1, P2, P3, P4)were selected, synthesized and coupled with KLH. Then the coupled polypeptides were used as antigens to immunize mice. After immunization, the mice serum was harvested, and the titer of the antibodies were determined with ELISA. The reactivity of antibodies to humans, rats, mice and NFM expressed in vitro were detected with Western blotting and immunohistochemistry(IHC). Results The four peptides had 山东大学学报 (医学版)56卷5期 -李昕,等.人神经丝中分子量亚单位多肽的免疫原性及其抗体的特异性 \=-strong immunogenicity, and antibodies of high titer were obtained. The IHC showed that all of the four antibodies were able to respond to NFM in human glioma, rat and mouse brain tissues. Western blotting indicated that anti-P1 and anti-P4 antibodies could react with the NFM in human glioma, rat and mouse brain tissues, and the anti-P1 antibody had two strips. Anti-P1, anti-P2 and anti-P3 could react with NFM expressed in vitro. Conclusion The four peptides have strong immunogenicity. The antibodies produced by mice can respond to NFM in human glioma, rat and mouse brain tissues and NFM expressed in vitro. They can be used in preparing McAb.

Key words: Middle subunit of neurofilament, Pepitide, Immunogenicity, Antibody specificity

中图分类号:

R392-33

李昕,刘建伟,薛皓,郑文慧,郑雯,秦泽明,温红玲,周晓莹,王志玉,赵丽. 人神经丝中分子量亚单位多肽的免疫原性及其抗体的特异性[J]. 山东大学学报 (医学版), 2018, 56(5): 58-63.

LI Xin, LIU Jianwei, XUE Hao, ZHENG Wenhui, ZHENG Wen, QIN Zeming, WEN Hongling, ZHOU Xiaoying, WANG Zhiyu, ZHAO Li. Human neurofilament medium polypeptide immunogenicity and specificity of antibodies[J]. Journal of Shandong University (Health Sciences), 2018, 56(5): 58-63.

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人神经丝中分子量亚单位多肽的免疫原性及其抗体的特异性

Human neurofilament medium polypeptide immunogenicity and specificity of antibodies

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