Separation of intact proteins by using polyhedral oligomeric silsesquioxane based hybrid monolithic capillary columns

被引：39

作者：

Liu, Zheyi ^{[1
]}

Ou, Junjie ^{[1
]}

Liu, Zhongshan ^{[1
,2
]}

Liu, Jing ^{[1
,2
]}

Lin, Hui ^{[1
,2
]}

Wang, Fangjun ^{[1
]}

Zou, Hanfa ^{[1
]}

机构：

[1] Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China

[2] Univ Chinese Acad Sci, Beijing 100049, Peoples R China

来源：

JOURNAL OF CHROMATOGRAPHY A | 2013年 / 1317卷

关键词：

Hybrid monolithic capillary column; Intact protein; Reversed phase liquid chromatography; High speed separation; LIQUID-CHROMATOGRAPHIC SEPARATION; SPECTROMETRY-BASED PROTEOMICS; ON-PLATE DIGESTION; MASS-SPECTROMETRY; TOP-DOWN; STATIONARY PHASES; PEAK-CAPACITY; ISOFORMS; IDENTIFICATION; PEPTIDES;

D O I：

10.1016/j.chroma.2013.09.004

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

High-efficient separation of intact proteins is still a huge challenge in proteome analysis of complex biological samples by using capillary columns. In this study, four FOSS-based hybrid monolithic capillary columns were prepared and applied in nano-flow liquid chromatography (Nano-LC) separation of intact proteins. It was observed that the POSS-based hybrid monolithic columns exhibit high permeability, good LC separation reproducibility and column efficiency for intact protein separation. The effects of different LC separation conditions such as flow rate, gradient steepness, column length and mobile phase additives on the LC separation efficiency of the POSS-based hybrid monolithic column were systematically examined. Finally, fast LC separation of 7 proteins mixture was realized in 2.5 min by using the optimized conditions on the 100 mu m i.d. POSS-based hybrid monolithic capillary column. (C) 2013 Published by Elsevier B.V.

引用

页码：138 / 147

页数：10

共 44 条

[11] Large-scale identification of proteins in human salivary proteome by liquid chromatography/mass spectrometry and two-dimensional gel electrophoresis-mass spectrometry [J].