A modified protein precipitation procedure for efficient removal of albumin from serum.
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 pre-fractionation 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.
Chen
Yi-Yun
YY
Core Facilities for Proteomic Research, Taipei, Taiwan.
Lin
Shu-Yu
SY
Yeh
Yuh-Ying
YY
Hsiao
He-Hsuan
HH
Wu
Chi-Yue
CY
Chen
Shui-Tsung
ST
Wang
Andrew H-J
AH
D000096:Acetone; D000818:Animals; D001797:Blood Protein Electrophoresis / Q000379:methods; D001798:Blood Proteins / Q000302:isolation & purification*; D005591:Chemical Fractionation; D011232:Chemical Precipitation; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008722:Methods; D051379:Mice; D040901:Proteomics / Q000379:methods*; D012709:Serum Albumin / Q000302:isolation & purification*; D014238:Trichloroacetic Acid
D001798:Blood Proteins; D012709:Serum Albumin; D000096:Acetone; D014238:Trichloroacetic Acid