[1]陈庆峰,马中良,王笑峰,等.抗粘蛋白1单链抗体的构建及其功能初步分析[J].陆军军医大学学报(原第三军医大学学报),2010,32(16):1770-1772.
 Chen Qingfeng,Ma Zhongliang,Wang Xiaofeng,et al.Construction and characterization of anti-mucin1 single chain Fv antibody fragment[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(16):1770-1772.
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抗粘蛋白1单链抗体的构建及其功能初步分析(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
32卷
期数:
2010年第16期
页码:
1770-1772
栏目:
论著
出版日期:
2010-08-30

文章信息/Info

Title:
Construction and characterization of anti-mucin1 single chain Fv antibody fragment
作者:
陈庆峰马中良王笑峰李福年郑玲莉
青岛大学医学院:附属医院乳腺外科,微生物教研室
Author(s):
Chen Qingfeng Ma Zhongliang Wang Xiaofeng Li Funian Zheng Lingli
Department of Breast Surgery, Affiliated Hospital, Department of Microbiology, Medical College, Qingdao University, Qingdao, Shandong Province, 266003, China
关键词:
乳腺肿瘤粘蛋白类单链抗体
Keywords:
breast neoplasms mucins single-chain antibody
分类号:
R392-33;R392.11
文献标志码:
A
摘要:
目的    分离鼠源抗粘蛋白1(mucin1,MUC1)单克隆抗体可变区基因,构建有结合功能的单链抗体,并初步检测结合活性。    方法    提取杂交瘤细胞总mRNA,合成cDNA后,PCR扩增鼠特异性抗体可变区编码基因,通过9个氨基酸的连接子构建表达单链抗体的原核重组表达载体,体外诱导表达纯化后,采用Western blot和流式细胞术方法分析单链抗体与变性或天然状态下抗原的结合活性。    结果    在杂交瘤细胞中成功分离到抗体轻重链可变区基因,重组链接后形成单链抗体能够在大肠杆菌中分泌表达,Western blot和流式细胞术分析表明该单链抗体能够与变性或天然状态下的T47D肿瘤细胞表面MUC1特异性结合。    结论    成功制备具有生物活性的抗肿瘤相关MUC1单链抗体,该单链抗体具有与MUC1特异性结合的生物学活性。
Abstract:

Objective To construct single chain variable fragment antibody (scFv) from a mouse monoclonal anti - mucin1 (MUC1) antibody secreting cell lines. Methods Isolation of mRNA of hybridoma cells, synthesis of cDNA, and antibody genotyping was performed using mouse antibody specific primers. Related variable genes were fused with a 9 - amino - acid linker to obtain a scFv,which was characterized on denatured and natural antigen by Western blotting and flow cytometry. Results Variable regions of antibodies heavy and light chain coding genes were isolated form a monoclonal anti - MUC1 hybridoma cell line. ScFv was obtained by the combination of the 2 variable genes, which could be produced in E.coli . Western blotting and FACS analysis showed that the obtained scFv could bind to nature and denatured MUC1 of T47D tumor cells specifically. Conclusion A functional anti - MUC1 scFv is obtained successfully, which can bind to tumour associated MUC1 specifically.

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更新日期/Last Update: 2010-08-31