基于纳升电喷雾-轨道阱超高分辨质谱对完整单克隆抗体药物进行分子量测定及糖型鉴定的条件探究 |
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基金项目:国家自然科学基金青年项目(21605085);中国科协青年人才托举工程项目(2017QNRC001) |
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中文摘要:整体分子量测定和糖型鉴定是抗体药物研发及生产中不可或缺的表征内容,而在完整蛋白层面进行测定可获得最为直接的测定结果。实现此类测定有赖于可提供较高分辨能力及质量精度的仪器;测定中涉及的相对复杂的参数设置可能受多种条件的干扰而产生误导性结果。该研究考察了利用纳升电喷雾离子源-轨道阱质谱仪以非变性质谱方式测定完整单克隆抗体药时,仪器分辨率、源内裂解、碰撞诱导碎裂、溶液组成等条件对单克隆抗体药物分子量及糖型测定结果的影响,以期为此类质谱分析提供有益的借鉴。 |
中文关键词:单克隆抗体 非变性质谱 完整蛋白 翻译后修饰 糖型 |
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Key Parameters for Molecular Weight Determination and Glycoform Identification on Intact Therapeutic Monoclonal Antibodies by Ultra high Resolution Orbitrap Mass Spectrometry with Nanospray Ionization |
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Abstract:Molecular weight determination and glycoform identification are the essential characterization data in stages ranged from development to quality control of therapeutic monoclonal antibodies(mAbs),while determinations on mAbs at the intact protein level have been demonstrated to provide straightforward and valid data,which require instruments with high resolving power and involve sophisticated parameter settings.To reduce possible artifacts,impacts of parameters were investigated,including nominal resolution,in source fragmentation,collision induced dissociation and solution composition on the molecular weight determination and glycoform identification of intact Abs by orbitrap mass spectrometry with nanospray ionization in native mass spectrometric mode,so as to provide a useful reference for this kind of mass spectrometric analysis. |
Key Words:monoclonal antibody native mass spectrometry intact protein post translational modification glycoform |
引用本文:代荣荣,马鑫,邵心阳,柴胡玲潇,王冠博.基于纳升电喷雾-轨道阱超高分辨质谱对完整单克隆抗体药物进行分子量测定及糖型鉴定的条件探究[J].分析测试学报,2020,39(1):122-127. |
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