本文選題：反相高效液相色譜法 + 重組類病毒顆粒疫苗��； 參考：《中國生物制品學(xué)雜志》2015年12期

【摘要】：目的反相高效液相色譜法(reverse phase-high performance liquid chromatography,RP-HPLC)檢測重組類病毒顆粒(virus-like particles,VLPs)疫苗原液中二硫蘇糖醇(DL-dithiothreitol,DTT)的殘留量,并對方法進(jìn)行驗證及初步應(yīng)用。方法按設(shè)定的色譜條件對檢測重組VLPs疫苗原液中DTT含量的RP-HPLC法進(jìn)行專屬性、精密性、準(zhǔn)確性驗證,確定方法的最佳線性范圍及最低檢測限,用該方法檢測3批重組HPV單價疫苗原液中的DTT殘留量。結(jié)果該方法檢測DTT時,不受溶液中溶劑和蛋白的干擾;線性范圍為1.5~15.0μg/ml(R~2=0.997),最低檢測限為1.5μg/ml;該方法檢測不同濃度DTT的平均相對標(biāo)準(zhǔn)偏差(relative standard deviation,RSD)均5%;重組HPV疫苗原液分別加入3、5、10μg/ml的DTT對照品溶液后,經(jīng)該方法檢測的回收率在80%~120%之間。3批次重組HPV單價疫苗原液中的DTT含量均小于1.5μg/ml。結(jié)論本實驗的RP-HPLC法專屬性、精密性良好,且準(zhǔn)確可靠,可用于重組HPV疫苗原液中DTT殘留量的檢測。
[Abstract]:Objective to determine the residue of DL-dithiothreitol (DL-dithiothreitol) in the original solution of recombinant viral particles (virus-like particlesus) vaccine by reverse phase-high performance liquid chromatography-RP-HPLC, and to verify the method and its preliminary application. Methods the RP-HPLC method for detecting DTT content in recombinant VLPs vaccine was determined by RP-HPLC according to the set chromatographic conditions. The specificity, accuracy and accuracy of the method were verified. The optimum linear range and the lowest detection limit of the method were determined. DTT residues in three batches of recombinant HPV monovalent vaccine were detected by this method. Results when DTT was detected by this method, it was not interfered by solvent and protein in solution. The linear range was 1.5 ~ 15.0 渭 g/ml (R ~ (2 +) 0.97) and the lowest detection limit was 1.5 渭 g 路ml ~ (-1). The recoveries of this method were between 80% and 120%. The DTT content in the original solution of recombinant HPV vaccine was less than 1.5 渭 g / ml. Conclusion the RP-HPLC method is specific, accurate and reliable, and can be used for the determination of DTT residues in recombinant HPV vaccine.
【作者單位】： 北京生物制品研究所有限責(zé)任公司;
【基金】：國家科技部重大新藥創(chuàng)制(2013ZX09402302)
【分類號】：O657.72;R392-33

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