基于直接注射-多级质谱全扫描技术的人血红蛋白快速肽图分析

引用本文: 杨湘芬, 张珂, 王世聪, 于娟, 牛晓雅, 陈蔚, 吴念, 宋青青, 宋月林. 基于直接注射-多级质谱全扫描技术的人血红蛋白快速肽图分析[J]. 分析化学, 2023, 51(3): 383-389. doi: 10.19756/j.issn.0253-3820.221384 shu

Citation: YANG Xiang-Fen, ZHANG Ke, WANG Shi-Cong, YU Juan, NIU Xiao-Ya, CHEN Wei, WU Nian, SONG Qing-Qing, SONG Yue-Lin. Rapid Tryptic Peptide Mapping of Human Hemoglobin Using Direct Infusion Coupled to MS/MS^ALL[J]. Chinese Journal of Analytical Chemistry, 2023, 51(3): 383-389. doi: 10.19756/j.issn.0253-3820.221384 shu

基于直接注射-多级质谱全扫描技术的人血红蛋白快速肽图分析

杨湘芬 ^1, ,
张珂 ^1, ,
王世聪 ^2, ,
于娟 ^2, ,
牛晓雅 ^1, ,
陈蔚 ^1, ,
吴念 ^1, ,
宋青青 ^{1,
,} ,
宋月林 ^{1,
,}

1.
北京中医药大学中药学院, 北京 102488;
2.
漳州片仔癀药业股份有限公司, 漳州 363000

通讯作者: 宋青青,E-mail:sqq1991qq@163.com; 宋月林,E-mail:syltwc2005@163.com

基金项目:
国家自然科学基金项目(Nos.81973444,82003911)资助。

摘要: 利用液相色谱-串联质谱法(LC-MS/MS)进行肽图分析,是目前单克隆抗体等蛋白类药物质量控制的主要方法,然而,LC-MS/MS分析具有费时、费溶剂和成本高等缺点。本研究以人血红蛋白(Humanhemoglobin,Hb)为例,考察利用直接注射-多级质谱全扫描技术(DI-MS/MS^ALL)进行肽图分析的可行性。Hb经胰蛋白酶水解后,多肽样品通过流动注射泵直接注入QTOF-MS离子源,采用MS/MS^ALL记录MS¹谱图以及每个单位质荷比窗口(1 Da)的MS²谱图信息,所获得的质谱信息与Skyline软件给出的21个理论肽段质谱信息进行匹配,初步指认了20个肽段。常规的LC-MS/MS肽图分析检测出了所有21个理论肽段。DI-MS/MS^ALL的MS¹图谱与LC-MS/MS的平均MS¹图谱相近,主要为各肽段带电荷数1~4的准分子离子信号,以双电荷为主,而MS²图谱均以y⁺离子为主。因此,DI-MS/MS^ALL分析蛋白肽图的能力与LC-MS/MS相似,并且具有分析通量高、溶剂用量少及成本低等优点,可作为快速分析肽图的有效工具,实现蛋白类药物的高通量分析。

关键词:

English

Rapid Tryptic Peptide Mapping of Human Hemoglobin Using Direct Infusion Coupled to MS/MS^ALL

1.
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China;
2.
Zhangzhou Pien-Tze-Huang Pharmaceutical Co., Ltd., Zhangzhou 363000, China

Corresponding author: SONG Qing-Qing, ; SONG Yue-Lin,

Received Date: 28 July 2022
Revised Date: 04 November 2022
Fund Project:
Supported by the National Natural Science Foundation of China (Nos. 81973444, 82003911).

Abstract: Peptide mapping currently serves as the primary means for quality control of protonic drugs through employing liquid chromatography-tandem mass spectrometry (LC-MS/MS) as the workhorse. Although being versatile, LC-MS/MS still suffers from shortcomings in terms of time-consuming, solvent-intensive and sophisticated instrumentation. Because of the extraordinary merit namely high throughput, the potential of direct infusion (DI) coupled to tandem mass spectrometry towards the tryptic peptide mapping was evaluated, and particularly, a new technique termed as MS/MS^ALL program was implemented to acquire MS² spectrum for each nominal m/z value, attributing to the involvement of the robust gas phase ion fractionation technique. Human hemoglobin (Hb) was utilized as a proof-of-concept target molecule. After thorough tryptic digestion, the peptide pool was directly injected into the ion source of mass spectrometer through an infusion pump, and MS/MS^ALL was programmed to record all desired MS² spectra, because the whole ion population was fractionated into sequential ion currents with 1 Da mass window. After matching the acquired MS¹ and MS² spectral information, 20 over 21 theoretical peptides that were suggested by Skyline software, were successfully captured, whilst all 21 peptides were detected by the well-defined LC-MS/MS program. Moreover, similar MS¹ spectra (average spectrum for LC-MS/MS) occurred between the two means, primarily including quasi-molecular ions bearing the charged states amongst 1-4. Either analytical method dominantly gave out characteristic y⁺ ions. Thereafter, comparable potential towards tryptic peptide mapping was yielded from DI-MS/MSALL in comparison with LC-MS/MS, and the DI-MS/MSALL exhibited many advantages including high throughput, solvent-saving and low-costing measurement. The results demonstrated that the DI-MS/MSALL could be an alternative analytical tool for LC-MS/MS.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

基于直接注射-多级质谱全扫描技术的人血红蛋白快速肽图分析

通讯作者: 宋青青,E-mail:sqq1991qq@163.com; 宋月林,E-mail:syltwc2005@163.com

English

Rapid Tryptic Peptide Mapping of Human Hemoglobin Using Direct Infusion Coupled to MS/MS^ALL

Corresponding author: SONG Qing-Qing, ; SONG Yue-Lin,

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于直接注射-多级质谱全扫描技术的人血红蛋白快速肽图分析

通讯作者: 宋青青,E-mail:sqq1991qq@163.com; 宋月林,E-mail:syltwc2005@163.com

English

Rapid Tryptic Peptide Mapping of Human Hemoglobin Using Direct Infusion Coupled to MS/MSALL

Corresponding author: SONG Qing-Qing, ; SONG Yue-Lin,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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