A simple affinity spin tube filter method for removing high-abundant common proteins or enriching low-abundant biomarkers for serum proteomic analysis.
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.
Wang
Young Y
YY
Department of Pathology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Cheng
Paul
P
Chan
Daniel W
DW
D015415:Biomarkers / Q000032:analysis*; D045744:Cell Line, Tumor; D002498:Centrifugation / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D011506:Proteins / Q000032:analysis*; D020543:Proteome; D040901:Proteomics / Q000379:methods*
D015415:Biomarkers; D007074:Immunoglobulin G; D011506:Proteins; D020543:Proteome