全柱成像毛细管等电聚焦电泳法测定4种生物制品等电点

目的：采用全柱成像毛细管等电聚焦电泳法（WCID-CIEF），测定多肽样品、胰岛素、融合蛋白疫苗和HPV病毒样颗粒的等电点。方法：采用50 mm×100 μm石英涂层毛细管。聚焦方法设置：1.5 kV聚焦1 min，3 kV聚焦4 min/8 min；检测波长：280 nm。结果：多肽样品、激素样品胰岛素和融合蛋白疫苗的等电点分别为10.98、5.87和5.86；病毒样颗粒样品HPV 31L1、HPV 33L1、HPV 45L1、HPV 52L1和HPV 58L1的实测等电点分别为8.17、7.86、7.77、7.91和7.69，并且实测等电点与理论等电点基本一致。全柱成像毛细管等电聚焦法测定生物制品专属性和精密度均符合验证要求。结论：该方法可用于生物制品的等电点分析。

Objective: To determine the isoelectric points of polypeptide, insulin, fusion protein vaccine and HPV virus-like particle(VLPs) by whole column imaging detection-capillary isoelectric focusing(WCIDCIEF). Methods: Cartridges of 50 mm×100 μm i. d. internally fluorocarbon-coated fused-silica capillary was used with the following settings:pre-focusing for 1 min under 1. 5 kV,followed by focusing at 3 kV for 4 min or 8 min;detection wavelength:280 nm. Results: The isoelectric points (pI) of the polypeptide,the hormone sample (insulin) and the fusion protein vaccine were 10. 98, 5. 87 and 5. 86, respectively. The five virus-like particles displayed pI values of 8. 17(HPV 31L1),7. 86(HPV 33L1),7. 77(HPV 45L1),7. 91(HPV 52L1) and 7. 69 (HPV 58L1). The obtained pI values were substantially consistent with the theoretical pI values. The specificity and precision of WCID-CIEF were validated. Conclusion: The method can be used for determing the isoelectric point of biological products.

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