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| 人CD123稳定表达细胞株的建立与鉴定 | |
| 引用本文： | 杨漾,黄亮,王娜,周剑峰. 人CD123稳定表达细胞株的建立与鉴定[J]. 实用全科医学, 2011, 9(5): 667-669,F0003 |
| 作者姓名： | 杨漾 黄亮 王娜 周剑峰 |
| 作者单位： | 华中科技大学同济医学院附属同济医院血液内科,武汉市,430030 |
| 基金项目： | 国家自然科学基金，国家科技重大专项课题 |
| 摘 要： | 目的探索建立人CD123稳定表达细胞株的方法,为进一步实现人CD123的抗体规模化生产提供材料。方法构建人CD123 CDS区真核表达载体（hCD123 CDS-pEGFP-N1）;经脂质体包裹转染中国仓鼠卵巢细胞CHO-S;经流式分选结合G418筛选,并用流式细胞术和Western印迹法检测人CD123蛋白的表达情况,初步筛选出人CD123高表达的CHO-S细胞株;并进一步用经人CD123 Fc重组融合蛋白免疫的小鼠血清检测该细胞株hCD123表达情况。结果测序及GenBank中序列比较结果显示扩增到的人CD123基因序列是正确的,酶切结果表明表达质粒构建正确;流式分选结合G418筛选得到1个人CD123高表达的细胞株——克隆8G9 CHO-S,其CD123-APC阳性细胞百分数为92.07%,用融合蛋白免疫的血清检测各细胞株人CD123表达情况,结果显示克隆8G9 CHO-S细胞和对照CHO-S细胞的CD123阳性率分别为（97.94±1.93）%和（2.84±1.35）%,差异具有统计学意义（P〈0.01）。结论建立了人CD123抗原稳定高表达的细胞株克隆8G9 CHO-S,为进一步实现人CD123的抗体规模化生产及其导向治疗奠定了坚实的基础。 |
| 关 键 词： | 人CDl23 稳定表达 CHO—S细胞株 细胞筛选 |
Establishment and Identification of a Cell Line Stably Expressing Hcd123 |
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| YANG Yang,HUANG Liang,WANG Na,et al.. Establishment and Identification of a Cell Line Stably Expressing Hcd123[J]. Applied Journal Of General Practice, 2011, 9(5): 667-669,F0003 | |
| Authors: | YANG Yang HUANG Liang WANG Na et al. |
| Affiliation: | YANG Yang,HUANG Liang,WANG Na,et al.Department of Hematology in Tongji Hospital,Tongji Medical College,Hua Zhong University of Science and Technology,Wuhan 430030,Hubei,China |
| Abstract: | Objective To establish a cell line that can stable express hCD123,which can be applied in the large-scale production of anti-CD123 antibody.Methods Expression vector of hCD123 CDS-pEGFP-N1 was constructed and transfected into chinese hamster ovarian cell line CHO-S using Lipofectamine 2000.G418 was used to select cell clones that could stably express human CD123.The expression of CD123 was verified by Western blot and flow cytometry.The cells that could highly express human CD123 were isolated by Flow cytom... |
| Keywords: | hCD123 Stable expression CHO-S cell line Cell screening |
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