蛋白质分离和鉴定的新技术新方法研究进展

杨开广; 张丽华; 张玉奎

doi:10.3969/j.issn.2095-476X.2012.05.001

Volume 27 Issue 5

September 2012

Article Contents

Abstract

References

YANG Kai-guang, ZHANG Li-hua and ZHANG Yu-kui. Recent advances of technique and method on the protein separation and identification in the proteomic studies[J]. Journal of Light Industry, 2012, 27(5): 1-7,12. doi: 10.3969/j.issn.2095-476X.2012.05.001

Citation: YANG Kai-guang, ZHANG Li-hua and ZHANG Yu-kui. Recent advances of technique and method on the protein separation and identification in the proteomic studies[J]. Journal of Light Industry, 2012, 27(5): 1-7,12. doi: 10.3969/j.issn.2095-476X.2012.05.001 shu

Recent advances of technique and method on the protein separation and identification in the proteomic studies

YANG Kai-guang ^1,2,3 ,
ZHANG Li-hua ^1,2,3 ,
ZHANG Yu-kui ^1,2,3,,

Dalian Inst. of Chem. Physics, Chinese Academy of Sci., Dalian 116023, China;
Key Lab. of Separation Sci. for Analytical Chemistry, Dalian 116023, China;
National Chromatographic R & A Center, Dalian 116023, China

Corresponding author: ZHANG Yu-kui,
Received Date: 2012-07-30
Available Online: 2012-09-15

Abstract

The new techniques and methods of protein separation and identification in the program "Novel Techniques and Methods for Protein Separation and Identification",supported by "National Basic Research Program of China",were reviewed.For high-abundance proteins depletion,strong cation exchange (SCX) chromatography was coupled with reversed phase liquid chromatography (RPLC) to develop two dimensional liquid chromatography,and high abundance proteins were applied as the templates to developed molecularly imprinted materials.For low abundance protein enrichment selectively,novel functional materials were developed for selectively capturing significant post-translated proteins,such as phosphopeptides/proteins and glycopeptides/proteins.For multidimensional,multi-mode and array protein separation,column switch recycling size exclusion chromatography (csrSEC) was coupled with RPLC to develop multidimensional liquid chromatography,and weak anion and cation exchange chromatography mixed-bed microcolumn were integrated with the immobilized trypsin reactor and RPLC-ESI/MS/MS to develop at high-throughput proteome platform.During the study of high-sensitive identification techniques,an online integrated platform for sample pretreatment,which was based on the hollow fiber membrane interface for solvent exchange,was established.New reagents were developed for the peptide derivatization,which decreased the limit of detection of peptide on the mass spectrum by 10-100 times.Core-shell nanoparticles were employed on the plate materials directly for the matrix-assisted laser desorption/ionization mass (MALDI).A predictive genetic algorithm was implemented for the optimization of filtering criteria to maximize the number of identified peptides for mass spectrum database searching.The new technology and new method for proteomic study provides an effective method.The development of selective adsorption materials provides a new way for high abundance protein depletion and low abundance proteins enrichment of proteomics research,and avoids the interference of the target protein identification in complex system.The platform of multidimensional and mode,array type of protein efficient separation technology as the core is constructed and realized the protein group of high efficiency,high flux,high reliability separation.With the new technology and new method development of high performance liquid chromatography/mass spectrometry as the core of high sensitive identification,the sensitivity and accuracy of low abundance protein mass spectrum identification are improved.
- proteome,
- high abundance protein depletion,
- low abundance protein enrichment,
- chromatography based separation,
- mass spectrum based identification
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