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| 抗体偶联小分子药物T-DM1的ELISA检测方法的建立 | |
| 引用本文： | 胡百卉,车津晶,许先兴,谌瑛,刘运龙,陈知航,程远国,许丽娜.抗体偶联小分子药物T-DM1的ELISA检测方法的建立[J].生物技术通讯,2014,25(5):693-696. |
| 作者姓名： | 胡百卉 车津晶 许先兴 谌瑛 刘运龙 陈知航 程远国 许丽娜 |
| 作者单位： | 1. 吉林农业大学动物科技学院,吉林长春,130062 2. 军事医学科学院微生物与流行病研究所,北京,100071 3. 第二炮兵总医院药学部,北京,100088 4. 吉林神话集团通化玉圣药业有限公司,吉林通化,134000 |
| 摘 要： | 目的:建立一种灵敏、特异、快速的ELISA方法,用于检测猕猴血清中T-DM1的含量。方法:采用双抗夹心ELISA法对T-DM1进行定量。以DM1单抗作为一抗包被到96孔板中,加入待检样品,之后再加入稀释好的猴血清吸附的羊抗人IgG-HRP(二抗),使之与结合到一抗上的T-DM1特异性结合,加底物显色,在酶标仪上读取D450nm值。结果:建立了检测T-DM1的ELISA方法并得到确证,方法的线性范围为0.625~80 ng/mL,定量下限为0.625 ng/mL,板内及板间精密度和准确度均在±15%以内,室温、冻融、稀释效应稳定性良好。结论:方法学验证表明ELISA法测定猴血清中T-DM1浓度的特异性、精密度和准确度均满足新生物制药临床前药代动力学研究指导原则要求,可用于T-DM1的检测。 |
| 关 键 词： | T-DM1 抗体偶联小分子药物 酶联免疫吸附法 药代动力学 人表皮生长因子受体2 |
Development of Enzyme Linked Immunosorbent Assay for Quantitative Determination of Antibody-Drug Conjugate T-DM1 |
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| Institution: | HU Bai-Hui, CHE Jin-Jing, XU Xian-Xing, CHEN Ying, LIU Yun-Long, CHEN Zhi-Hang, CHENG Yuan-Guo, XU Li-Na( 1. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130062; 2. Institute of Mi- crobiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071; 3. Department of Pharmacy, Second Artillery General Hospital, Beijing 100088; 4. Jilin Tonghua Yusheng Myth Grou9 Pharmaceutical Co., Ltd, Tonghua 134000; China) |
| Abstract: | Objective： To develop a sensitive, specific and rapid ELISA method for detection of T-DM1 in rhesus monkey serum. Methods： A quantitative sandwich ELISA for T-DM1 quantified was developed using DM1 monoclonal antibody for capturing and goat anti-human IgG-HRP for detecting. Following that, color was devel- oped by the substrate solution and the reaction was stopped by stop solution. Then the plate was analyzed with a microplate reader at a wavelength of 450 nm. Results： An ELISA assay was developed with a wide dynamic range of concentrations from 0.625 to 80 ng/mL with a lowest quantification of 0.625 ng/mL, both accuracy of the intra- and inter-assay were less than 15%. Meanwhile, well stability of T-DM1 was shown by evaluation on different storage conditions and dilution with serum. Conclusion： This method is highly sensitive, accurate, specific, and reproducibility, which was proven to be a feasible quantitative method for the detection of T-DM1. |
| Keywords: | T-DM1 antibody-drug conjugate ELISA pharmacokinetics human epidermalgrowth factor receptor-2 |
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