目的 建立13价肺炎球菌结合疫苗解吸附处理总蛋白含量测定方法并验证。方法 13价肺炎球菌结合疫苗经解吸附处理，利用Lowry法对其总蛋白含量进行测定，对建立的方法进行线性、准确度、重复性及耐用性验证。结果 适宜的解吸附条件为加入10%体积的0.3 mol/L NaOH。在蛋白标准品40~200 μg/mL范围内，标准曲线线性良好，决定系数>0.995； 20、40、60、80、120 μg/mL的蛋白标准品回收率在95.25%~104.77%之间，方法准确度良好；样品在不同时间重复测定6次的实验结果相对标准偏差为3.82%，方法重复性良好；解吸附后室温下分别放置15和30 min，测定结果之间差异无统计学意义（t值分别为0.059和0.238，P值>0.05），且测定结果的相对标准偏差分别为4.82%和5.14%，方法耐用性良好。结论 建立的方法可有效、准确、稳定地检测13价肺炎球菌结合疫苗中总蛋白含量。

Objective To establish and validate a method for determination of total protein content after desorption in 13-valent pneumococcal conjugate vaccine.Methods The total protein content of the 13-valent pneumococcal conjugate vaccine was determined by the Lowry method after desorption. The linearity, accuracy, repeatability and durability of the established method were verified.Results The optimum desorption condition was to add 10% volume of 0.3 mol/L NaOH. Within the range of 40-200 μg/mL of protein standard, the linearity of the standard curve was good, coefficient of determination＞0.995. In the accuracy test, the recovery rates of protein standards at concentrations of 20,40,60,80 and 120 μg/mL ranged from 95.25% to 104.77%,demonstrating good accuracy. In the repeatability test,the relative standard deviation (RSD) of 6 test results of the sample at different time points was 3.82%, indicating good repeatability.In the durability test, no statistically significant difference (t = 0.059 and 0.238, P > 0.05) was observed in test results after samples were stored at room temperature for 15 or 30 min post-desorption, with RSD values of 4.82% and 5.14%，respectively, indicating good durability.Conclusion The established method can effectively, accurately and stably detect the total protein content of 13-valent pneumococcal conjugate vaccine.