Extensive deuterium back-exchange in certain immobilized pepsin columns used for H/D exchange mass spectrometry

被引：35

作者：

Wu, Y ^{[1
]}

Kaveti, S ^{[1
]}

Engen, JR ^{[1
]}

机构：

[1] Univ New Mexico, Dept Chem, Albuquerque, NM 87131 USA

来源：

ANALYTICAL CHEMISTRY | 2006年 / 78卷 / 05期

关键词：

D O I：

10.1021/ac0518497

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Pepsin digestion prior to mass analysis increases the spatial resolution of hydrogen exchange mass spectrometry experiments. Online digestion with immobilized pepsin is advantageous for several reasons including better digestion efficiency. We have found that certain immobilized pepsin columns cause substantial deuterium back-exchange, rendering the data unusable. When pepsin immobilized on a POROS support was used for online digestion, back-exchange was within the expected range and was similar to the back-exchange of deuterated peptides produced by in-solution pepsin digestion. However, when pepsin immobilized onto selected polystyrene-divinylbenzene supports was used for online digestion with the same system, deuterium loss was extremely high. The effect seems linked to the properties of the solid support used to conjugate the pepsin.

引用

页码：1719 / 1723

页数：5

共 17 条

[1] Use of different proteases working in acidic conditions to improve sequence coverage and resolution in hydrogen/deuterium exchange of large proteins [J].