Simultaneous quantification of major capsid protein of human papillomavirus 16 and human papillomavirus 18 in multivalent human papillomavirus vaccines by liquid chromatography-tandem mass spectrometry
单位：[a]Beijing Key Laboratory for Precancerous Lesion of Digestive Disease, Department of Gastroenterology, Beijing Friendship Hospital, National Clinical Research Center for Digestive Disease, Beijing Digestive Disease Center, Capital Medical University, Beijing, China临床科室国家中心消化分中心消化内科首都医科大学附属北京友谊医院[b]Division of HIV/AIDS and Sexually Transmitted Virus Vaccines, National Institutes for Food and Drug Control (NIFDC), No.31 Huatuo Street, Daxing District, Beijing.102629, China[c]Division of Physics and Chemistry, National Institutes for Food and Drug Control (NIFDC), No.2 Tiantanxili, Dongcheng District, Beijing 10 0 050, China[d]Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Canada
Human papillomavirus (HPV) vaccination is the most effective mean to prevent HPV infection and cervical carcinoma. Licensed HPV prophylactic vaccines are formulated to contain a defined amount of different major capsid protein (L1), the critical antigen to elicit protection. No method is currently available to simultaneously quantify individual L1s in multivalent vaccines, presenting a daunting challenge for the quality control of HPV vaccines. Here, HPV16 and HPV18 L1 can be analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using tryptic digestion without pre-digestion reduction and alkylation in multiple reaction monitoring (MRM) mode. Two signature peptides were selected to be the markers of the two L1s and can be well separated within 5.1 min. Their linear calibration curves were both obtained in the range of 20-500 nmol/L (R-2 > 0.990). To HPV16 L1, intra/inter assay precisions and accuracies of the assay were below 11% and between 83.96-113.57%. While for HPV18 L1, they were below 12% and between 81.40-103.49%. In addition, the limits of quantitation (LOQ) were as low as 2.8 nmol/L for HPV16 L1 and 1.7 nmol/L for HPV18 L1, respectively, representing about 68 and 112 times more sensitive than those obtained with Smith Bicinchoninic Acid (BCA) assay. This LC-MS/MS method can be applied to the quantification of both bulk products and the final multivalent vaccines. This method is superior to the current assays in terms of sensitivity, specificity, precision, accuracy and throughput; it could become the method of choice for absolute quantification of proteins in multivalent vaccines. (C) 2020 Elsevier B.V. All rights reserved.
基金：
National Science and Technology Major Projects of Drug Discovery [2018ZX09101001]
语种：
外文
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出版当年[2019]版：
大类|2 区化学
小类|2 区生化研究方法2 区分析化学
最新[2025]版：
大类|2 区化学
小类|1 区生化研究方法2 区分析化学
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出版当年[2018]版：
Q1CHEMISTRY, ANALYTICALQ1BIOCHEMICAL RESEARCH METHODS
最新[2023]版：
Q1BIOCHEMICAL RESEARCH METHODSQ1CHEMISTRY, ANALYTICAL
第一作者单位：[a]Beijing Key Laboratory for Precancerous Lesion of Digestive Disease, Department of Gastroenterology, Beijing Friendship Hospital, National Clinical Research Center for Digestive Disease, Beijing Digestive Disease Center, Capital Medical University, Beijing, China[b]Division of HIV/AIDS and Sexually Transmitted Virus Vaccines, National Institutes for Food and Drug Control (NIFDC), No.31 Huatuo Street, Daxing District, Beijing.102629, China
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推荐引用方式(GB/T 7714)：
Tingting Ning,Shanshan Sun,Jianhui Nie,et al.Simultaneous quantification of major capsid protein of human papillomavirus 16 and human papillomavirus 18 in multivalent human papillomavirus vaccines by liquid chromatography-tandem mass spectrometry[J].JOURNAL of CHROMATOGRAPHY a.2020,1619:doi:10.1016/j.chroma.2020.460962.
APA：
Tingting Ning,Shanshan Sun,Jianhui Nie,Mengyi li,Weijin Huang...&Youchun Wang.(2020).Simultaneous quantification of major capsid protein of human papillomavirus 16 and human papillomavirus 18 in multivalent human papillomavirus vaccines by liquid chromatography-tandem mass spectrometry.JOURNAL of CHROMATOGRAPHY a,1619,
MLA：
Tingting Ning,et al."Simultaneous quantification of major capsid protein of human papillomavirus 16 and human papillomavirus 18 in multivalent human papillomavirus vaccines by liquid chromatography-tandem mass spectrometry".JOURNAL of CHROMATOGRAPHY a 1619.(2020)
化学