全柱成像毛细管等电聚焦电泳技术在重组蛋白疫苗电荷异质性分析中的应用

Application of whole-column imaging capillary isoelectric focusing electrophoresis in charge heterogeneity analysis of recombinant protein vaccines

重组蛋白分子表征的稳定性是保证疫苗发挥免疫原性及抗原性的关键。重组蛋白疫苗与治疗性蛋白制品一样,因体外合成和纯化过程的复杂性而表现出异质性。其中,电荷异质性是重组蛋白质的关键质量属性之一。电荷异质性监测有助于推动生产工艺的优化,反映产品生产的一致性和稳定性等,对保证重组蛋白产品的安全和质量至关重要。开发可靠快速、灵敏稳健的电荷异质性分析方法一直是生物制药行业所追求的目标。经典的电荷异质性分析方法主要有离子交换色谱(IEC)、毛细管区带电泳(CZE)以及全柱成像毛细管等电聚焦电泳(WCID-CIEF)等,其中WCID-CIEF提供的等电点(pI)和峰形的生化指纹有助于对疫苗多蛋白成分的分析。基于以上该文将对WCID-CIEF的原理和特点及其在重组蛋白疫苗质量控制中的应用进行概述。

Recombinant protein vaccine is a kind of therapeutic protein,the recombinant protein products usually exhibit heterogeneity due to the complex in vitro synthesis and purification processes.Charge heterogeneity is a critical quality attributes of recombinant proteins.Charge heterogeneity monitoring helps to promote the optimization of the production process and reflect the consistency and stability of product production,which is essential to ensure the safety and quality of recombinant protein products.Developing a reliable,high-specificity,high-sensitivity and robust analysis method for analyzing charge heterogeneity has always been the goal pursued by the biopharmaceutical industry.Classical methods for analyzing charge heterogeneity mainly include ion exchange chromatography(IEC),whole-column imaging capillary isoelectric focusing electrophoresis(WCID-CIEF)and capillary zone electrophoresis(CZE).WCID-CIEF is a reliable and high-sensitivity analytical method,the isoelectric point(pI)values and peak provided by WCID-CIEF can be used as biochemical finger prints to facilitate the analysis of various protein components in vaccines.Therefore,the principle and characteristics of WCID-CIEF and the application of WCID-CIEF in quality control of existing recombinant protein vaccines will be introduced in this review.