A modified protein precipitation procedure for efficient removal of albumin from serum

Yi Yun Chen, Shu Yu Lin, Yuh Ying Yeh, He Hsuan Hsiao, Chi Yue Wu, Shui Tsung Chen, Andrew H.J. Wang

Research output: Contribution to journalArticlepeer-review

149 Citations (Scopus)

Abstract

Proteomic analysis of sera and the quest for identifying serum proteins as disease markers have often been hampered by the predominance of several highly abundant proteins including albumin and immunoglobulins. Prior albumin depletion so as to enrich for otherwise undetectable serum components is therefore a prerequisite in mining the serum proteome. In the course of evaluating several available methods and commercial kits, we have been able to refine the albumin depletion protocols and establish a modified albumin removal method using trichloroacetic acid (TCA)/acetone. Changes in major protein bands were monitored by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1-D SDS-PAGE) and used as the first screening strategy to evaluate and optimize for the precipitation experimental conditions. Our method showed better performance in efficiency, specificity, and costs in comparison with two commercially available albumin removal kits, and provides a simple prefractionation step for the proteomic analysis of serum biomarkers. Albumin isolated by the modified method is in the native state. Our method may offer a rapid method for purifying serum albumin in large scale.

Original languageEnglish
Pages (from-to)2117-2127
Number of pages11
JournalElectrophoresis
Volume26
Issue number11
DOIs
Publication statusPublished - Jun 2005
Externally publishedYes

Keywords

  • Albumin removal
  • Serum proteins
  • Trichloroacetic acid precipitation

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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