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单链抗体骨架1区在与抗原结合中的作用

Role of framework 1 of single chain variable fragment in binding to antigen
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摘要 [目的 ]探讨单链抗体可变区骨架 1区对抗体特异性的影响 .[方法 ]应用分子克隆技术 ,以分离自重症肌无力患者胸腺的抗原结合片段Fab 6 37为亲本 ,构建了完整序列的单链抗体ScFv 6 37以及在重链和轻链可变区骨架 1区N端各缺失 8个氨基酸的单链抗体ScFv6 37Δ8aa ,在大肠杆菌表达后 ,应用放射免疫测定法检查单链抗体与抗原 人乙酰胆碱受体的结合活性 .[结果 ]单链抗体ScFv 6 37Δ8aa不能与人乙酰胆碱受体结合 ,然而在同一试验中 ,单链抗体ScFv 6 37能高亲合性地与人乙酰胆碱受体结合 .[结论 ]单链抗体骨架 1区的结构可影响抗体与抗原表位的结合特异性 . OBJECTIVE To investigate the influences of framework 1 of single chain variable fragments (ScFv) to their specificity.?METHODS?The intact ScFv?637 and ScFv?637 with the deletion of first 8 amino acids in framework 1 regions of both heavy and light chains (ScFv?637?Δ8aa) were constructed from their parental antigen binding fragment (Fab), derived from thymus of a patient with myasthenia gravis by using molecular cloning technique. After expressed in E coli, the binding activities of ScFv?637 and ScFv?637?Δ8aa to human acetylcholine (hAChR) receptor were determined by radioimmunoassay.?RESULTS?ScFv?637?Δ8aa was unable to bind to hAChR, whereas ScFv?637 had a high affinity to bind to hAChR in the same experiment.?CONCLUSION?The structure of framework 1 region of ScFv affects the binding specificity of antibody to epitope of antigen.
作者 杨康鹃 孟繁平 Stassen M. de Baets M.
机构地区 延边大学医学院细胞生物学与医学遗传学教研室 延边大学医学院微生物学与免疫学教研室 马斯特里赫特大学医学院神经学系
出处 《延边大学医学学报》 CAS 2003年第2期86-89,共4页 Journal of Medical Science Yanbian University
基金 国家自然科学基金 (3996 0 0 72 ) 教育部高等学校骨干教师资助计划基金
关键词 单链抗体 骨架1区 抗体特异性 分子克隆 抗原-人乙酰胆碱受体 结合活性 antibodies, monoclonal antibody specificity framework
分类号 R392.11 [医药卫生—免疫学]
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参考文献11

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二级参考文献8

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  • 8杨康鹃,孟繁平,YGraus,MdeBaets.重组质粒对大肠杆菌 DH5_α 的转化[J].延边大学医学学报,1997,20(2):69-70. 被引量：3

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延边大学医学学报
2003年 第2期

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