全柱成像毛细管等电聚焦电泳分析重组蛋白质药物电荷异质性

Imaged Capillary Isoelectric Focusing analysis of charge heterogeneity of recombinant protein drugs

关键词:  全柱成像毛细管等电聚焦电泳  电荷异质性  重组蛋白质药物  单抗  融合蛋白  德谷胰岛素

Key words:  imaged capillary isoelectric focusing  charge heterogeneity  recombinant protein drugs  secukinumab  fusion protein  insulin degludec

摘要:

目的 分析德谷胰岛素、苏金单抗、阿柏西普和重组人生长激素4种重组蛋白质药物的电荷异质性，为其质量检控提供可靠且快速的分析方法。方法 利用iCE3全柱成像毛细管等电聚焦电泳(imaged capillary isoelectric focusing，iCIEF)分析仪对待测重组蛋白质药物进行检测，将待测样品与载体两性电解质按一定比例混合，在1 500~3 000 V聚焦条件下进行检测。结果 德谷胰岛素等电聚焦电泳图为单峰，等电点平均值为5.25。苏金单抗共检测到4个等电点，其中等电点8.66为主峰，占总峰面积的67.78%。阿柏西普共检测到12个等电点，有6个等电点的峰面积占总峰面积>10%，RSD均<0.10%。重组人生长激素共有3个等电点，其中等电点5.45为主峰占总峰面积的87.60%。结论 利用iCIEF技术能够有效检测德谷胰岛素、苏金单抗、阿柏西普和重组人生长激素的电荷异质性。该方法对4种蛋白质药物的质量检控具有重要意义。

Abstract:

OBJECTIVE To analyze the charge heterogeneity of insulin degludec, sujinzumab, aflibercept and recombinant human growth hormone, and to provide a reliable and rapid analysis method for quality control of four recombinant protein drugs. METHODS The recombinant protein drugs to be tested were detected with imaged capillary isoelectric focusing(iCIEF). The samples were mixed with carrier amphoteric electrolyte in a certain proportion and detected under the focusing conditions of 1 500~3 000 V. RESULTS The isoelectric focal electrophoresis of insulin degludec showed a single peak with an average isoelectric point of 5.25. Four isoelectric points were detected of secukinumab, which 8.66 was the main peak, accounting for 67.78% of the total peak area. A total of 12 isoelectric points were detected for aflibercept fusion protein, and the peak area of 6 isoelectric points accounted for >10% of the total peak area, and the RSD was <0.10%. rhGH had 3 isoelectric points, of which 5.45 was the main peak, accounting for 87.60% of the total peak area. CONCLUSION iCIEF technique is useful to detect the charge heterogeneity of insulin degludec, sujinzumab, aflibercept and rhGH. It is of great significance for quality control for four protein drugs.