Pepsin immobilized on high-strength hybrid particles for continuous flow online digestion at 10,000 psi.
Pepsin immobilized on high-strength hybrid particles for continuous flow online digestion at 10,000 psi.
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DOI：
10.1021/ac301749h
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发表时间：
2012-08-21
影响因子：
7.4
通讯作者：
Engen, John R.
中科院分区：
文献类型：
--
作者：
Ahn, Joomi;Jung, Moon Chul;Wyndham, Kevin;Yu, Ying Qing;Engen, John R.
将胃酶固定在乙桥杂化(BEH)颗粒上，并以完全在线的形式评估消化性能，目的是将该颗粒用于氢-氘交换质谱仪(HDX-MS)实验。由于BEH粒子的机械强度很强，它们可以在10,000磅/平方英寸的压力下承受长期持续的高压。在线消解是在恒压条件下进行的，具有连续的溶剂流动，与其他方法不同的是，压力或流动是循环的。正如预期的那样，10,000磅/平方英寸的消化效率提高了，与1,000磅/平方英寸的消化相比，可重复产生更多的肽多肽。用BEH胃酶颗粒制成的原型色谱柱表现出较好的性能，与其他固定化胃酶颗粒的重离子背交换性能相似。这些颗粒可以很容易地合并到现有的HDX MS工作流程中，以便在需要快速、高效和可重复的在线胃酶消化的实验中提供更多的多肽覆盖。
Pepsin was immobilized on ethyl-bridged hybrid (BEH) particles and digestion performance was evaluated in a completely online format, with the specific intent of using the particles for hydrogen deuterium exchange mass spectrometry (HDX MS) experiments. Because the BEH particles are mechanically strong, they could withstand prolonged, continuous high-pressure at 10,000 psi. Online digestion was performed under isobaric conditions with continuous solvent flow, in contrast to other approaches where the pressure or flow is cycled. As expected, digestion efficiency at 10,000 psi was increased and reproducibly produced more peptic peptides versus digestion at 1,000 psi. Prototype columns made with the BEH pepsin particles exhibited robust performance and deuterium back-exchange was similar to that of other immobilized pepsin particles. These particles can be easily incorporated in existing HDX MS workflows to provide more peptide coverage in experiments where fast, efficient, and reproducible online pepsin digestion is desired.
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影响因子：
3.4
作者：
Kim, Byoung Chan;Lopez-Ferrer, Daniel;Lee, Sang-Mok;Ahn, Hye-Kyung;Nair, Sujith;Kim, Seong H.;Kim, Beom Soo;Petritis, Konstantinos;Camp, David G.;Grate, Jay W.;Smith, Richard D.;Koo, Yoon-Mo;Gu, Man Bock;Kim, Jungbae
通讯作者：
Kim, Jungbae
影响因子：
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通讯作者：
Geromanos, SJ
影响因子：
2
作者：
Munoz, Marina;de Ancos, Begona;Pilar Cano, M.
通讯作者：
Pilar Cano, M.
影响因子：
4.4
作者：
Lopez-Ferrer, Daniel;Petritis, Konstantinos;Hixson, Kim K.;Heibeck, Tyler H.;Moore, Ronald J.;Belov, Mikhail E.;Camp, David G., II;Smith, Richard D.
通讯作者：
Smith, Richard D.
影响因子：
3.4
作者：
Kim, Jungbae;Kim, Byoung Chan;Lopez-Ferrer, Daniel;Petritis, Konstantinos;Smith, Richard D.
通讯作者：
Smith, Richard D.