| 应用ELISA测定睫状神经营养因子中卡那霉素残留量 |
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| 引用本文： | 王兰,毕华,饶春明.应用ELISA测定睫状神经营养因子中卡那霉素残留量[J].药物分析杂志,2011(6). |
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| 作者姓名： | 王兰 毕华 饶春明 |
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| 作者单位： | 中国药品生物制品检定所重组技术产品室; |
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| 基金项目： | 国家科技重大专项课题(课题编号:2009ZX09307-001) |
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| 摘 要： | 目的:建立ELISA检测卡那霉素残留量的方法并进行方法学验证,用于重组技术产品的质量控制。方法:采用间接竞争ELISA方法,样本中残留的卡那霉素将和微孔板上预包被的偶联抗原竞争抗卡那霉素抗体,加入酶标二抗后,用3,3,5,5-四甲基联苯胺(TMB)底物显色,样本吸光值与残留卡那霉素的含量成负相关,用酶标仪进行检测并用SOFT MAX分析软件进行分析。结果:ELISA方法测定卡那霉素残留量在0.5~40.5 ng.mL-1范围内线性良好,且符合四参数方程式:y=(A-D)/1+(X/C)B]+D,r2≥0.99;板内变异小于15%,板间变异小于20%,实验重复性好,加标回收率在80%~120%之间;该方法检测与庆大霉素、盐酸四环素、链霉素和氨苄西林药物未显示交叉反应。应用该法对1批睫状神经营养因子(CNTF)原液的卡那霉素残留量进行测定,结果表明,每人用剂量CNTF(100μg)卡那霉素残留量小于0.5 ng。结论:该方法具有灵敏度高、操作简便、检测迅速等特点,可用于重组技术产品中卡那霉素残留量的检定。
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| 关 键 词： | ELISA 卡那霉素 残留量 重组生物制品 质量控制 |
Determination of residual kanamycin in ciliary neurotrophic factor(CNTF) by ELISA |
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| WANG Lan,BI Hua,RAO Chun-ming.Determination of residual kanamycin in ciliary neurotrophic factor(CNTF) by ELISA[J].Chinese Journal of Pharmaceutical Analysis,2011(6). |
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| Authors: | WANG Lan BI Hua RAO Chun-ming |
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| Institution: | WANG Lan,BI Hua,RAO Chun-ming(Division of Biopharmaceuticals,National Institute for the Control of Pharmaceutical and Biological Products,Beijing 100050,China) |
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| Abstract: | Objective:To establish ELISA method for determination of residual kanamycin for the quality control of recombinant products.Method:Using indirect competitive ELISA,residual kanamycin in the test sample can compete with pre-coated antigen in the microplate for antibody specific for kanamycin.An enzyme-linked antibody specific for kanamycin was added to the wells,then a substrate solution was added to the wells and color developed in negative proportion to the amount of residual kanamycin.The results were rea... |
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| Keywords: | ELISA residual kanamycin recombinant biological product quality control |
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