Internally calibrated quantification of VEGF in human plasma by fluorescence immunoassays in disposable elastomeric microfluidic devices

Herein we report on a proof of principle for the reproducible quantification of Vascular Endothelial Growth Factor (VEGF) in human plasma by fluorescence sandwich immunoassays using disposable polydimethylsiloxane (PDMS) microfluidic chips. The system requires 100 times less sample than typical clinical blood tests, while its current quantification limit is established at 4 pM. The in-built calibration method of spiking the plasma with known concentrations of commercially available antigen avoids common sources of error and improves the reliability of the test results. The demonstrated technique is important for immunoassay applications in fundamental scientific research and “point-of-care” (POC) biomedical diagnostics. In particular, the system is immediately applicable to microfluidic quantification of VEGF in human plasma in cancer studies.