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

被引:85
作者
Wang, YY
Cheng, P
Chan, DW
机构
[1] Johns Hopkins Med Inst, Dept Pathol, Baltimore, MD 21287 USA
[2] Johns Hopkins Singapore, Tumor Marker Lab, Singapore, Singapore
关键词
biomarker; serum; spin tube filter; two-dimensional gel electrophoresis;
D O I
10.1002/pmic.200390036
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:243 / 248
页数:6
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