重组人粒细胞巨噬细胞集落刺激因子宿主DNA残留的研究Detection of Residual DNA in rhGM-CSF
林秀菁;孔雯雯;
LIN Xiu-jing,KONG Wen-wen(Guangzhou Baiyunshan Baidi Bio-Technology Co.,Ltd,Guangzhou,Guangdong 511495,China)
摘要(Abstract)：
目的建立及优化重组人粒细胞巨噬细胞集落刺激因子(rhGM-CSF)宿主DNA残留的检测方法。方法基于固相斑点杂交原理,以rhGM-CSF的工程菌DNA为模板,制备地高辛标记的探针,用试剂盒及药典的方法处理阳性对照品,并将其与探针进行杂交,采用底物显色法和化学发光法显色,建立灵敏度较高的检测方法;运用建立的方法检测2批次rhGM-CSF原液中DNA残留量。结果样品经试剂盒处理,杂交后采用化学发光法显色灵敏度较高,可达到10 pg;2批rhGM-CSF原液每人份剂量的宿主DNA残留量均小于100 pg。结论该方法具有检测灵敏度高、准确性好、操作简便等优势,可用于rhGM-CSF原液的检测及质量控制。
Objective To establish and optimize the method for testing the residual DNA in rhGM-CSF.Methods The total DNA of engineering bacteria was labeled by digoxin as probe,and positive DNA was prepared by DNA purification kit and traditional method to hybridize with the probe,then chemiluminescent and chromogenic substrate method were used for developing process.Based on the established method,the residual DNA in 2 batchs of bulk products of rhGM-CSF was assayed.Results The higher sensitive assayed that included the positive DNA processed using kit and chemiluminescent method showed 10 pg DNA.The residual DNA in 2 batches of bulk products was less than 100 pg per dosage for one person.Conclusion This method is sensitive,accurate and convenient,which can be used for the quality control of the bulk products of rhGM-CSF.
关键词(KeyWords)：
重组人粒细胞巨噬细胞集落刺激因子;DNA残留;地高辛;杂交
rhGM-CSF;residual DNA;digoxin;hybridization
基金项目(Foundation):
作者(Authors):
林秀菁;孔雯雯;
LIN Xiu-jing,KONG Wen-wen(Guangzhou Baiyunshan Baidi Bio-Technology Co.,Ltd,Guangzhou,Guangdong 511495,China)
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