Identification of low-abundance proteins via fractionation of the urine proteome with weak anion exchange chromatography

Proteome Science

Ying-Chin Ko⁴  Chun-Hsiung Huang⁷  Jeff Yi-Fu Chen⁶  Jiing-Chuan Chen³  Jue-Liang Hsu²  Cheng-Chi Chen¹  Yu-Jen Wu¹  Chih-Ming Lu⁵ 

[1] Department of Beauty Science, Meiho University, Pingtung, Taiwan;Graduate Institute of Biotechnology, National Pingtung University of Science and Technology, Pingtung, Taiwan;Department of Food Science and Nutrition, Meiho University, Pingtung, Taiwan;Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan;Department of Urology, Buddhist Da Lin Tzu Chi General Hospital, Chiayi, Taiwan;Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, Taiwan;Department of Urology, Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

关键词: Urine;    Proteomic;    Fractionation;    DEAE-Sephacel;    Weak anion exchange chromatography;   

Others  :  824927
DOI  :  10.1186/1477-5956-9-17

 received in 2010-12-05, accepted in 2011-04-08,  发布年份 2011

【 摘 要 】

Background

Low-abundance proteins are difficultly observed on the two-dimensional gel electrophoresis (2-DE) maps of urine proteome, because they are usually obscured by high-abundance proteins such as albumin and immunoglobulin. In this study, a novel fractionation method was developed for enriching low-abundance proteins by removing high-abundance proteins and progressive elution with salts of various concentrations.

Results

Stepwise weak anion exchange (WAX) chromatography, which applied DEAE-Sephacel resin with non-fixed volume elution, was used to fractionate urine proteome prior to performing 2-DE. Urine proteome was separated into four fractions by progressively eluting the column with 0 M, 50 mM, 100 mM, and 1 M NaCl solutions. Most of the heavy and light immunoglobulin chains appeared in the eluent. After the high-abundance proteins were removed, various low-abundance proteins were enriched and could be easily identified. The potential of this method for obtaining diversified fractionations was demonstrated by eluting the column separately with Na₂SO₄ and MgCl₂ solutions. The 2-DE maps of the fractions eluted with these different salt solutions of identical ionic strength revealed markedly different stain patterns.

Conclusion

The present study demonstrated that this fractionation method could be applied for purposes of enriching low-abundance proteins and obtaining diversified fractionations of urine, and potentially other proteomes.

【 授权许可】

   
2011 Lu et al; licensee BioMed Central Ltd.

【 图 表 】

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