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。

Objective　To validate the Lowry method 2 for the determination of pertussis toxin (PT) , filamentous hamagglutinin (FHA)，and pertactin (Prn) contents in pertussis antigen. Methods　PT, FHA, and Prn national standards were diluted and the standard curve was established in the concentration range of 0-50 μg/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. Results　The 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. Conclusion　The 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.