结核DNA疫苗宿主蛋白残留检测方法的建立  

Establishment of Detection Method for Host Protein Residues in Tuberculosis DNA Vaccine

作　　者：谭奇凤 李婷 孔雯雯 倪世明 黄明 梁艳[2] 吴雪琼[2] 郭润姿 TAN Qi-feng;LI Ting;KONG Wen-wen;NI Shi-ming;HUANG Ming;LIANG Yan;WU Xue-qiong;GUO Run-zi(Guangzhou BaiYunShan BaiDi Biotechnology CO.LTD.Guangzhou 511495,China;Tuberculosis Key Laboratory,the 8th Medical Center of PLA General Hospital,Beijing 100091,China;Gu′an Dingtai Haigui Biotechnology CO.LTD,Langfang 065500,China)

机构地区：[1]广州白云山拜迪生物医药有限公司,广东广州511495 [2]解放军总医院第八医学中心结核病重点实验室,北京100091 [3]固安鼎泰海规生物科技有限公司,河北廊坊065500

出　　处：《海峡药学》2022年第12期45-48,共4页Strait Pharmaceutical Journal

基　　金：国家科技重大专项资助(艾滋病和病毒肝炎等重大传染病的防治,2018ZX10731301-005-003)。

摘　　要：目的建立结核DNA疫苗中大肠杆菌宿主蛋白残留量检测的方法。方法采用双抗夹心酶联免疫吸附法(ELISA法)建立结核DNA疫苗中宿主蛋白残留量检测的方法,对方法的准确度、专属性、线性范围和耐用性等进行验证,并采用建立的方法对结核DNA疫苗原液进行测定。结果大肠杆菌宿主蛋白含量在2~24ng·L^(-1)范围内,相关系数R 2>0.990,线性关系良好;该法宿主蛋白含量在高中低浓度范围内,回收率在70%~120%之内,准确度良好;该法不易受辅料、核酸、孵育温度和孵育时间影响,专属性和耐用性较好;三批原液的宿主蛋白残留量(HCP)均<1μg·mg^(-1)。结论此方法有较好的准确度、专属性、重复性、中间精密度和耐用性,可用于结核DNA疫苗的宿主蛋白残留量的检测。OBJECTIVE To develop a detection method for the residual content of E.coli host protein in tuberculosis DNA vaccine.METHODS The residual content of host protein of tuberculosis DNA vaccine was detected by double antibody sandwich enzyme-linked immunosorbent assay(ELISA),and the accuracy,specificity,linear range and durability of the method were verified.The established method was used to test the TB DNA vaccine stock solution.RESULTS The linear relationship was good which that the correlation coefficient R 2 was>0.990 and the content of E.coli host protein was in the range of 2-24 ng·L^(-1).The accuracy recovery was 70%-120%with the content of host protein was in the range of high,middle and low concentrations.The method not only has strong specificity which that is less affected by excipients and nucleic acids,but also the durability is good which that is less affected by changing the incubation temperature and incubation time.The residual amount of host protein(HCP)of three batches of stock solution is less than 1μg·mg^(-1).CONCLUSION This method has good accuracy,specificity,repeatability,intermediate precision and durability,and can be used to determine the residual content of host protein in Tuberculosis DNA vaccine.

关 键 词：宿主蛋白含量 结核DNA疫苗 ELISA法 检测 

分 类 号：R297[医药卫生—民族医学]