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  • Title: [Using size-exclusion chromatography to quantify the 146S antigen in inactivated foot-and-mouth disease vaccine].
    Author: Xu Y, Zou X, Li C, Zhu Y, He T, Yang Y, Lin X, Song Y, Zheng J, Zhang S, Zhao Q.
    Journal: Sheng Wu Gong Cheng Xue Bao; 2018 May 25; 34(5):676-684. PubMed ID: 29893075.
    Abstract:
    The aim of this study is to quantify the 146S antigen in foot-and-mouth disease virus (FMDV) inactivated vaccine by size-exclusion chromatography (SEC). The analysis was performed on a TSKgel G4000SWXL column (7.8 mm×30 cm), with a pH 7.2 buffer salt system as the mobile phase. The flow rate was 0.6 mL/min, the injection volume was 100 μL and the detection wavelength was 259 nm. The calibration curve was established by using purified inactivated FMDV (serotype O) 146S antigen; 3 batches of vaccine formulated by inactivated antigen solution were tested to verify the accuracy, reproducibility, specificity and tolerability of the method. At last 16 batches of vaccine were determined by the SEC method. Results showed a good linearity between peak area and concentration of 146S antigen in the range between 0.56 and 67.42 μg/mL (R2=0.996, n=10), and the average recovery rate of 146S antigen in the 3 batches of vaccine formulated in lab were 93.6% (RSD=2.7%, n=3), 102.3% (RSD=2.6%, n=3), and 95.5% (RSD=5.1%, n=3). The method was proved accurate and reliable with good reproducibility (RSD=0.5%, n=6), and applied to determine 16 batches of the commercial FMDV vaccine. According to the above results, the SEC method is high effective for 146S antigen quantify in the inactivated FMDV vaccine and would provide strong support for the vaccine quality control. 旨在应用体积排阻色谱法测定口蹄疫灭活疫苗中的146S 抗原含量。使用TSKgel G4000SWXL (7.8 mm×30 cm)色谱柱，以pH 7.2 的缓冲盐体系作为流动相，流速为0.6 mL/min，进样量为100 μL，检测波长为259 nm。以口蹄疫病毒 (O 型) 灭活146S 抗原纯化样品建立标准曲线；使用灭活抗原液配制3 份口蹄疫灭活疫苗，并进行精密度、重复性、特异性、耐受性验证；应用该方法快速测定16 批疫苗的146S 含量。结果表明，抗原浓度在0.56–67.42 μg/mL范围内，其峰面积与浓度的线性关系良好 (R2=0.996，n=10)，3 份疫苗146S 抗原测定回收率分别为93.6% (RSD=2.7%，n=3)、102.3% (RSD=2.6%，n=3)、95.5% (RSD=5.1%，n=3)，方法重复性好、准确性强 (RSD=0.5%，n=6)，且操作简便、高效，对16 批疫苗的测定结果较理想。应用该方法有望快速高效地检测口蹄疫灭活疫苗的146S 抗原含量，为疫苗的质量控制提供有力支持。.
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