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刊物信息

期刊名称:药物分析杂志
主管单位:中国科学技术协会
主办单位:中国药学会
承办:中国食品药品检定研究院
主编:金少鸿
地址:北京天坛西里2号
邮政编码:100050
电话:010-67012819,67058427
电子邮箱:ywfx@nicpbp.org.cn
国际标准刊号:ISSN 0254-1793
国内统一刊号:CN 11-2224/R
邮发代号:2-237
 

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牛痘病毒粒细胞巨噬细胞集落刺激因子注射液的生物学活性测定方法研究

Development of bioassays for vaccinia virus/GM-CSF injection

分类号:R917
出版年·卷·期(页码):2020,40 (1):52-57
DOI: 10.16155/j.0254-1793.2017.01.01
-----摘要:-------------------------------------------------------------------------------------------

目的: 建立牛痘病毒粒细胞巨噬细胞集落刺激因子(vaccinia virus/GM-CSF)注射液的生物学活性测定方法。方法: 采用噬斑法进行感染滴度测定,结合定量聚合酶链反应(quantitative polymerase chain reaction,Q-PCR)进行病毒基因组滴度/感染滴度测定;采用TF-1细胞增殖法,四参数方程拟合处理试验数据,检测不同浓度梯度的样品感染Hela细胞后表达的GM-CSF生物学活性;采用ELISA法,二次方程拟合处理试验数据,检测不同浓度梯度样品感染Hela细胞后的GM-CSF的表达量;采用ELISA法进行β半乳糖苷酶活性测定;采用肿瘤细胞(UACC 257)和正常细胞(GM 00038)进行溶瘤活性测定。结果: 测定了1批牛痘病毒粒细胞巨噬细胞集落刺激因子注射液,其病毒感染滴度为6.5×108 PFU·mL-1,病毒基因组滴度/感染滴度为27,各浓度梯度孔GM-CSF表达量均大于10 pg,GM-CSF生物学活性为181 rGCU,溶瘤活性为144 rOnU,β-半乳糖苷酶活性为124 rBU。结论: 对牛痘病毒GM-CSF注射液的生物学活性进行了相对全面的测定,各个项目均符合规定,能够用于该产品的质量控制,同时对同类产品的质量控制具有借鉴意义。

-----英文摘要:---------------------------------------------------------------------------------------

Objective: To establish methods for biological activity determination of vaccinia virus/granulocyte macrophage colony stimulating factor(GM-CSF)injection.Methods: Infectious titer was measured by plaque assay and viral genome titer/infectious titer was measured by a combination of quantitative polymerase chain reaction (Q-PCR)and plaque assay.The biological activity of GM-CSF was detected by TF-1 cell proliferation assay and curve fitting was performed by four-parameter equation.The expression of GM-CSF in Hela cells infected by different concentrations of virus was detected by ELISA method and curve fitting was performed by quadratic equation.The activity of β-galactosidase was determined by ELISA. The oncolytic activity was determined by the tumor cells(UACC 257) and normal cells(GM 00038).Results: A batch of vaccinia virus/GM-CSF injection was determined.The viral infectious titer was 6.5×108 PFU·mL-1,viral genome titer/infectious titer was 27,GM-CSF expression level was more than 10 pg in all gradient wells,biological activity of GM-CSF was 181 rGCU,oncolytic activity was 144 rOnU,and β-galactosidase activity was 124 rBU. Conclusion: The biological activity of vaccinia virus/GM-CSF injection has been determined in a relatively comprehensive way,and all the items meet the requirements,which can be used for the quality control of related products,and also provide significance reference for the quality control of similar products.

-----参考文献:---------------------------------------------------------------------------------------

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