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Статья
Quench me if you can: Alpha-2-macroglobulin trypsin complexes enable serum biomarker analysis by MALDI mass spectrometry
2021
One of the main functions of alpha-2-macroglobulin (A2M) in human blood serum is the binding of all classes of protease. It is known that trypsin, after such interaction, possesses modified proteolytic activity. Trypsin first hydrolyzes two bonds in A2M's ‘bait region’, and the peptide VGFYESDVMGR is released from A2M. In this work, specifics of the A2M-trypsin interaction were used to determine A2M concentration directly in human blood serum using MALDI mass-spectrometry. Following exogenous addition of trypsin to human blood serum in vitro, the concentration of the VGFYESDVMGR peptide was measured, using its isotopically-labeled analogue ( O), and A2M concentration was calculated. The optimized mass spectrometric approach was verified using a standard method for A2M concentration determination (ELISA) and the relevant statistical analysis methods. It was also shown that trypsin's modified proteolytic activity in the presence of serum A2M can be used to analyze other serum proteins, including potential biomarkers of pathological processes. Thus, this work describes a promising approach to serum biomarker analysis that can be technically extended in several useful directions. 705 715 18
Цитирование
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Версии
- 1. Version of Record от 2021-06-01
Метаданные
Название журнала
- Biochimie
Том
- 185
Страницы
- 87-95
Финансирующая организация
- Russian Science Foundation
Номер гранта
- 075-15-2020-934
Тип документа
- journal article
Тип лицензии Creative Commons
- CC BY
Правовой статус документа
- Свободная лицензия
Источник
- scopus
