| 重组人心肌肌钙蛋白I单克隆抗体的制备及鉴定 |
| |
| 引用本文： | 佟晓丽,张春明,王德芝. 重组人心肌肌钙蛋白I单克隆抗体的制备及鉴定[J]. 生物医学工程与临床, 2009, 13(2). DOI: 10.3969/j.issn.1009-7090.2009.03.019 |
| |
| 作者姓名： | 佟晓丽 张春明 王德芝 |
| |
| 作者单位： | 中国医学科学院,中国协和医科大学放射医学研究所,天津分子核医学重点实验室,天津,300192 |
| |
| 摘 要： | 目的 制备重组人心肌肌钙蛋白I(rhcTnI)的单克隆抗体,并对其单克隆抗体的特性进行鉴定.方法 利用纯化后的rhcTnI作为免疫原,常规免疫Balb/c小鼠,取其脾细胞与同系骨髓瘤细胞Sp2/0进行常规融合,用甲基纤维素半固体培养基法获得单克隆抗体杂交瘤细胞株,利用ELISA、Western blot等方法对单克隆抗体的特性进行鉴定.结果 筛选出3株稳定分泌抗rhcTnI的单克隆抗体杂交瘤细胞株,分别命名为4B3、7D5和3E6.3株单克隆抗体的免疫球蛋白亚类分别为IgG1(4B3、7D5)和IgG2b(3E6).3株单克隆抗体与rhcTnI发生特异性结合,而与大肠杆菌菌体蛋白、乳酸脱氢酶(LDH)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)无特异性交叉反应,3株单克隆抗体的亲和力常数分别为1.76×109Umol、143×109L/mol和1.04×109L/mol.结论 获得了效价高、特异性好的抗rhcTn I的单克隆抗体,为cTnI诊断试剂盒的研制奠定了基础.
|
| 关 键 词： | 人心肌肌钙蛋白I单克隆抗体 甲基纤维素半固体培养基 |
Preparation and identification of monoclonal antibodies against recombinant human cardiac troponin I |
| |
| TONG Xiao-li,ZHANG Chun-ming,WANG De-zhi. Preparation and identification of monoclonal antibodies against recombinant human cardiac troponin I[J]. Biomedical Engineering and Clinical Medicine, 2009, 13(2). DOI: 10.3969/j.issn.1009-7090.2009.03.019 |
| |
| Authors: | TONG Xiao-li ZHANG Chun-ming WANG De-zhi |
| |
| Affiliation: | Tianjin Key Laboratory of Molecular Nuclear Medicine;Institute of Radiation Medicine;Chinese Academy of Medical Sciences and Peking Union Medical College;Tianjin 300192;China |
| |
| Abstract: | Objective To obtain monoclonal antibodies(mAb) against recombinant human cardiac troponin I(rhcTn I) and identify their characteristics. Methods Balb/c mice were immunized with rhcTn I. The spleen cells of the immunized mice were fused with Sp2/0 cells by regular cell fusion and hybridoma cell lines were screened by methyl cellulose semi-solid culture medium contained HAT. The characteristics of these monoclonal antibodies were determined by the techniques of ELISA, Western blot etc. Results Three hybridoma... |
| |
| Keywords: | human cardiac troponin I monoclonal antibodies methyl cellulose semi-solid culture medium |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
