Detecting Residual Host Genomic DNA in Biotech Drugs by Real-Time Quantitative PCR

doi:10.3969/j.issn.2095-2341.2014.02.12

Curr. Biotech. ›› 2014, Vol. 4 ›› Issue (2): 142-145.DOI: 10.3969/j.issn.2095-2341.2014.02.12

Detecting Residual Host Genomic DNA in Biotech Drugs by Real-Time Quantitative PCR

CAO Chen-hua, LIU Xiao-zhi, DUAN Yue-jiao, LIU Su-xia, ZHAO Wei, CHANG Liang, JI Li-man, GAO Jian

State Key Laboratory of Antibody Research and Development; New Drug Research and Development Company Ltd., North China Pharmaceutical Corporation, Shijiazhuang 050015, China

Received:2013-12-13 Online:2014-03-25 Published:2014-03-03

Abstract

Abstract: We detected residual host DNA in yeast expressing biotech drugs by SYBR GreenⅠ fluorescent based real-time quantitative polymerase chain reaction. The detection sensitivity of this method was up to 1.0 fg/μL. The linear correlation coefficient of standard curve was above 0.99 when DNA concentration was bwtween 1.0 fg/μL to 1.0 ng/μL. Three batch of samples was determined, and the residual host DNA were 8.635×105 fg/μL、6.265×102 fg/μL and 1.436 fg/μL respectively. The results suggested that this easy-handled, rapid and highly sensitive method could be used for the determination of residual yeast DNA in biotech drug products.

Key words: real-time quantitative PCR, residual host genomic DNA, biotech drugs, quality control

CAO Chen-hua§, LIU Xiao-zhi§, DUAN Yue-jiao, LIU Su-xia, ZHAO Wei, CHANG Liang, JI Li-man*, GAO Jian*. Detecting Residual Host Genomic DNA in Biotech Drugs by Real-Time Quantitative PCR[J]. Curr. Biotech., 2014, 4(2): 142-145.

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https://swjsjz.magtechjournal.com/EN/Y2014/V4/I2/142

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Detecting Residual Host Genomic DNA in Biotech Drugs by Real-Time Quantitative PCR

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