| 抗rhNDPK-A单克隆抗体的制备及鉴定 |
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| 引用本文： | 李雪玲,王一飞,熊盛,张美英,李冰,李久香,王红,郑佐娅,王从朱.抗rhNDPK-A单克隆抗体的制备及鉴定[J].细胞与分子免疫学杂志,2004,20(1):86-88. |
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| 作者姓名： | 李雪玲 王一飞 熊盛 张美英 李冰 李久香 王红 郑佐娅 王从朱 |
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| 作者单位： | 1. 暨南大学生物医药研究开发基地,广东,广州,510632
2. 上海第二医科大学医学检验重点实验室,上海,200023 |
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| 基金项目： | 国家高技术研究发展计划 (863)资助 (No .2 0 0 1AA2 1 50 4 1 )，广东省科委重大攻关项目资助 (No.A1 0 90 2 0 7) |
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| 摘 要： | 目的 :研制抗rhNDPK A (recombinanthumannucleosidediphosphatekinase A)单克隆抗体 (mAb) ,并鉴定其特性。 方法 :以纯化的rhNDPK A免疫BALB/c小鼠 ,采用杂交瘤技术制备抗rhNDPK AmAb ;用免疫双扩散鉴定Ig亚类 ;West ernblot鉴定mAb的特异性 ;间接ELISA检测mAb的腹水效价、亲和常数 ,并进行表位分析。结果 :获得 6株可分泌特异性mAb的抗rhNDPK A的杂交瘤细胞系 2D9、8C7、13E2、15D9、15E3和 2 0D9,Ig亚类均为IgG1;其效价为 1× 10 -4～5× 10 -6;亲和常数为 4 .5× 10 -9～ 2 .8× 10 -10 mol/L ;共有3个抗原表位。结论 :获得抗rhNDPK A的mAb ,为进一步用于临床诊断和实验研究创造了条件
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| 关 键 词： | rhNDPK-A 单克隆抗体 特性鉴定 |
| 文章编号： | 1007-8738(2004)01-0086-03 |
| 修稿时间： | 2003年4月30日 |
Preparation and characterization of monoclonal antibodies against recombinant human NDPK-A |
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| LI Xue-ling ,WANG Yi-fei ,XIONG Sheng ,ZHANG Mei-ying ,LI Bing ,LI Jiu-xiang ,WANG Hong ,ZHENG Zuo-ya ,WANG Cong-zhu Biomedicine Research & Development Center of Jinan University,Guangzhou , Shanghai Municipal Key Laboratory of Medical Technique,Shanghai Second Medical University,Shanghai ,China.Preparation and characterization of monoclonal antibodies against recombinant human NDPK-A[J].Journal of Cellular and Molecular Immunology,2004,20(1):86-88. |
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| Authors: | LI Xue-ling WANG Yi-fei XIONG Sheng ZHANG Mei-ying LI Bing LI Jiu-xiang WANG Hong ZHENG Zuo-ya WANG Cong-zhu Biomedicine Research & Development Center of Jinan University Guangzhou Shanghai Municipal Key Laboratory of Medical Technique Shanghai Second Medical University Shanghai China |
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| Institution: | Biomedicine Research & Development Center of Jinan University, Guangzhou 510632, China. |
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| Abstract: | AIM: To prepare monoclonal antibodies (mAb) against recombinant human nucleoside diphosphate kinase-A(NDPK-A) and characterize their properties. METHODS: BALB/c mice were immunized with rhNDPK-A, and mAb was prepared by hybridoma technique. Ig subclass and specificity was analysed by double immunodiffusion and western blot respectively. The titres of mAbs in ascitic fluid, relative affinity and epitopes recognized by mAbs were determined by indirect ELISA. RESULTS: 6 hybridoma cell lines secreting anti-rhNDPK-A mAbs were obtained. Their Ig subclass belonged to IgG1. The titers of 6 mAbs in ascitic fluid were 1x10(-4)-5x10(-6). Relative affinity of mAbs were 4.5x10(-9)-2.8x10(-10) mol/L. They recognized 3 different epitopes on rhNDPK-A molecule. CONCLUSION: 6 mAbs against rhNDPK-A have been prepared successfully which provide useful reagent for clinical diagnosis and further research. |
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| Keywords: | rhNDPK-A monoclonal antibody characterization |
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