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目的:建立毛细管电泳法测定重组人乳头瘤病毒原液的纯度。方法:采用50μm×57 cm(有效长度50 cm)无涂层毛细管。进样时方法设置为:进样5 kV,20 s;分离电压为15 kV,维持30 min,两端加压138 kPa;温度:25℃;检测波长:220 nm。结果:毛细管电泳法测定重组人乳头瘤病毒(HPV)16型、18型原液主峰分离度分别为2.4和3.0,HPV 16L1和HPV 18L1原液在浓度为0.125～0.750 mg.mL-1时线性关系良好(HPV 16L1,r=0.9836;HPV 18L1,r=0.9931)。HPV 16L1和HPV 18L1原液的精密度和稳定性试验的RSD均<2.0%。结论:本文建立的方法可用于重组人乳头瘤病毒原液的纯度检测。
Objective: To establish a capillary electrophoresis method to determine the purity of recombinant human papillomavirus (HPV) stock solution. Methods: 50μm × 57 cm (effective length 50 cm) uncoated capillaries. The injection method was set to: injection 5 kV, 20 s; separation voltage of 15 kV for 30 min, both ends of the pressure 138 kPa; temperature: 25 ℃; detection wavelength: 220 nm. Results: The main peaks of recombinant human papillomavirus (HPV) 16 and 18 were 2.4 and 3.0, respectively. Capillary electrophoresis showed that the linearity of HPV16L1 and HPV 18L1 was good at 0.125-0.750 mg.mL-1 (HPV 16L1, r = 0.9836; HPV 18L1, r = 0.9931). The RSDs for the precision and stability tests of both HPV 16L1 and HPV 18L1 stock solutions were <2.0%. Conclusion: The method established in this paper can be used for the purity of recombinant human papillomavirus (HPV) stock solution.