A simple affinity spin tube filter method for removing high-abundant common proteins or enriching low-abundant biomarkers for serum proteomic analysis

Proteomics. 2003 Mar;3(3):243-8. doi: 10.1002/pmic.200390036.

Abstract

Although it is possible to identify new proteins from crude cell extracts using proteomics technology, it is often difficult to elucidate low-abundant biomarkers in the presence of a large amount of high-abundant proteins in serum. We have developed a simple and rapid method using an affinity spin tube filter to remove high-abundant common proteins and enrich the low-abundant biomarkers. The affinity spin tube filter contains protein G, coupled with antibodies against either high-abundant proteins or specific proteins of interest. After incubating with serum, the flow-through or the elute was collected and analyzed by two-dimensional gel electrophoresis. By using this affinity spin tube filter, the possibilities of identifying new biomarkers are shown. This technique could be used for large-scale sample preparation for high-throughput proteomic analysis.

MeSH terms

  • Biomarkers / analysis*
  • Cell Line, Tumor
  • Centrifugation / methods
  • Electrophoresis, Gel, Two-Dimensional
  • Humans
  • Immunoglobulin G / chemistry
  • Proteins / analysis*
  • Proteome
  • Proteomics / methods*

Substances

  • Biomarkers
  • Immunoglobulin G
  • Proteins
  • Proteome