Plasmonic alloy has attracted much interest in tailoring localized surface plasmon resonance (LSPR) for recent biosensing techniques. In particular, pap er-based plasmonic substrates allow capillary-driven lateral flow as well as three-dimensional metal nanostructures, and therefore they become actively transferred to LSPR-based biosensing such as surface-enhanced Raman spectroscopy (SERS) or metal-enhanced fluorescence (MEF). However, employing plasmonic alloy nanoislands on heat-sensitive substrate is still challenging, which significantly inhibits broad-range tailoring of the plasmon resonance wavelength (PRW) for superior sensitivity. Here we report paper-based plasmonic substrate with plasmonic alloy of Au/Ag nanocomposites for highly sensitive MEF and SERS biosensing applications. The nanofabrication procedures include concurrent deposition of Au and Ag below 100 degrees C without any damage on cellulose fibers. The Au/Ag nanocomposites feature nanoplasmonic alloy with single plasmon peak as well as broad-range tunability of PRW by composition control. This paper-based plasmonic alloy substrate enables about twofold enhancement of fluorescence signals and selective MEF after paper chromatography. The experimental results clearly demonstrate extraordinary enhancement in SERS signals for picomolar detection of folic acid as a cancer biomarker. This new method provides huge opportunities for fabricating plasmonic alloy on heat-sensitive substrate and biosensing applications.