Herein we report on reliable reproducible quantification of protein analytes in human serum by fluorescence sandwich immunoassays in disposable PDMS microfluidic chips. The system requires 1000 times less sample than typical clinical blood tests and is specifically shown to measure ferritin down to 250 pM in human serum. The in-built calibration method of spiking the serum with known concentrations of commercially available antigen avoids common sources of error and improves the reliability of the test results. The reported microfluidic system is an important new tool for fundamental scientific research, offering sensitive immunoassay measurements in small but complex biosamples. The system is also a further step towards comprehensive affordable "point-of-care" biomedical diagnostics.