期刊
JOURNAL OF PROTEOME RESEARCH
卷 12, 期 12, 页码 5996-6003出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr400877e
关键词
immunoprecipitation; MRM; plasma biomarkers; biomarker verification; colon cancer
资金
- National Cancer Institute through the Clinical Proteomic Technology Assessment for Cancer (CPTAC) program [5U24CA126479]
Quantitative analysis of protein biomarkers in plasma is by the availability of high-quality antibodies. An alternative multiple reaction monitoring mass spectrometry (IP-MRM). We compared IP-MRM to ELISA for the analysis of six colon cancer biomarker candidates (metalloproteinase inhibitor 1 (TIMP1), cartilage oligomeric matrix protein (COMP), thrombospondin-2 (THBS2), endoglin (ENG), mesothelin (MSLN) and matrix metalloproteinase-9 (MMP9)) in plasma from colon cancer patients and noncancer controls. Proteins were analyzed by multiplex immunoprecipitation from plasma with the ELISA capture antibodies, further purified by SDS-PAGE, digested and analyzed by stable isotope dilution MRM. IPMRM provided linear responses (r = 0.978-0.995) between 10 and 640 ng/mL for the target proteins spiked into a mock plasma matrix consisting of 60 mg/mL bovine serum albumin. Measurement variation (coefficient of variation at the limit of detection) for IP-MRM assays ranged from 2.3 to 19%, which was similar to variation for ELISAs of the same samples. IP-MRM and ELISA measurements for all target proteins except ENG were highly correlated (r = 0.67-0.97). IP-MRM with high-quality capture antibodies thus provides an effective alternative method to ELISA for protein quantitation in biological fluids.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据