| Abstract
| - Screening of mutant mice for subtle phenotypes requiressensitive, high-throughput analyses of sentinel proteinsin functional pathways. The cytokine TNF-α is upregulatedduring inflammatory reactions associated with autoimmune diseases. We have developed a method to monitor the concentration of TNF-α under physiological conditions. TNF-α is captured, purified, and concentrated usingmonoclonal antibody-coated microbeads. The capture isefficient (>80%) and can be used in the concentrationrange <100 pg/mL to >50 ng/mL, as determined bydetection of 125I-labeled TNF-α. The bead capture ofTNF-α can be combined with direct detection by MALDI-MS for sample concentrations of >10 ng/mL. TNF-α canbe captured and detected from diluted mouse serum, withminimal interferences observed in the MALDI spectrum.This method is adaptable to high-throughput samplehandling with microfluidic devices and automated massspectrometric analysis.
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