Lowry法检测百日咳抗原中百日咳毒素、丝状血凝素、黏附素含量方法验证  

Validation of Lowry method for pertusis toxin,filamentous hemagglutinin,and pertactin contents detection in pertussis antigen

作　　者：邢盛宇 邓海清[1] 季庚辰 张艳红[1] 张钰东 王宇 吴晗 尹珊珊[1] Xing Shengyu;Deng Haiqing;Ji Gengchen;Zhang Yanhong;Zhang Yudong;Wang Yu;Wu Han;Yin Shanshan(R&D Center,Beijing Minhai Biotechnology Co.,Ltd.,Beijing 102600,China)

机构地区：[1]北京民海生物科技有限公司研发中心,北京102600

出　　处：《国际生物制品学杂志》2024年第3期151-155,共5页International Journal of Biologicals

摘　　要：目的对Lowry法2检测百日咳抗原中百日咳毒素(pertussis toxin, PT)、丝状血凝素(filamentous hemagglutinin, FHA)、黏附素(pertactin, Prn)含量进行方法学验证。方法将PT、FHA、Prn国家标准品系列稀释, 在0~50 μg/ml浓度范围内建立标准曲线。在已知浓度的PT、FHA、Prn精制液中加入国家标准品配制加标样品, 采用Lowry法2检测蛋白含量, 考察准确度和精密度。对原液生产中使用的400 mmol/L PBS进行专属性试验。分别在检测后10、20、30 min再进行检测以考察方法的耐用性。结果回收率介于91.1%~110.7%;准确度相对标准偏差为1.8%~9.2%, 批间精密度相对标准偏差为2.6%~6.2%;400 mmol/L PBS对样品测定无干扰;检测样品在30 min内读取数据有效。结论该法具有良好的线性、准确性、精密性、专属性和耐用性, 可用于百日咳抗原中PT、FHA、Prn含量的测定。ObjectiveTo validate the Lowry method 2 for the determination of pertussis toxin(PT),filamentous hamagglutinin(FHA),and pertactin(Prn)contents in pertussis antigen.MethodsPT,FHA,and Prn national standards were diluted and the standard curve was established in the concentration range of 0-50μg/ml.PT,FHA,and Prn refined solutions were added with national standards to prepare standard samples,and were tested for protein contents by Lowry method 2.The specificity was tested for 400 mmol/L PBS used in bulk production.Robustness was validated by testing samples 10,20,and 30 min after primary test.ResultsThe recoveries ranged from 91.1%to 110.7%,the accuracy relative standard deviations(RSDs)ranged from 1.8%to 9.2%,and the intermediate precision RSDs ranged from 2.6%to 6.2%.The 400 mmol/L PBS had no interference on the determination of samples.Test results were reliable within 30 min.ConclusionThe Lowry method 2 has good linearity,accuracy,precision,specificity and robustness,and can be used for the determination of PT,FHA,and Prn contents in pertussis antigen.

关 键 词：Lowry法 百日咳菌苗 蛋白检测 方法学验证 

分 类 号：R927.2[医药卫生—药学]