UHPLC-LTQ-Orbitrap-MS分析人血白蛋白及人免疫球蛋白中非目标蛋白成分

    Analysis of Non-target Proteins in Human Albumin and Human Immunoglobulin by UHPLC-LTQ- Orbitrap-MS

    • 摘要: 目的 采用超高效液相色谱-串联线性离子阱-轨道阱组合高分辨质谱技术对人血白蛋白及人免疫球蛋白中非目标蛋白成分进行定性分析。方法 采用ACQUITY UPLC peptide BEH C18(300Å,1.7 μm,2.1 mm×100 mm)色谱柱,以0.1%甲酸水溶液-0.1%甲酸乙腈为流动相进行梯度洗脱,质谱采用Full MS/dd-MS2(TopN)扫描模式。结果 从人血白蛋白及人免疫球蛋白中共鉴定出非目标蛋白52个,其中人血白蛋白样本中鉴定非目标蛋白25个,人免疫球蛋白样本中鉴定非目标蛋白27个。结论 建立的定性方法可快速、准确、系统地识别出人血白蛋白及人免疫球蛋白中多种蛋白,为该类制剂质量控制及进一步的临床应用提供参考。

       

      Abstract: OBJECTIVE To analyze and identify non-target proteins in human albumin and human immunoglobulin by ultra high performance liquid chromatography tandem linear ion trap orbitrap mass spectrometry. METHODS The extract was separated on a ACQUITY UPLC peptide BEH C18(300Å, 1.7 μm, 2.1 mm×100 mm) column and the gradient elution was performed with mobile phase consisting of 0.1% formic acid aqueous solution-0.1% formic acid acetonitrile. The analytes were detected in Full MS/dd-MS2(TopN). RESULTS A total of 52 non-target proteins were identified from human albumin and human immunoglobulin. Among them, 25 non-target proteins were identified in human albumin samples, and 27 non-target proteins were identified in human immunoglobulin samples. CONCLUSION The established qualitative method can rapidly, accurately and systematically identify various proteins in human albumin and human immunoglobulin. The results provide reference for the quality control of the preparation as well as its further clinical application.

       

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