This paper demonstrates a microfluidic chip for multiple immunoassays on electrospun (ES) nanofibrous membranes made of PVDF. This microchip enables the detection of multiple pairs of protein-protein interactions in one experiment by crossing parallel arrays of microchannels. This chip uses ESPVDF membrane with high specific surface area as the substrate to adsorb protein for immobilized immunoassays so that increased levels of protein adsorption may lower the LOD in immunoassays. We employ a simple and effective method for chip assembly by scotch tape to incorporate the ESPVDF membrane into the microchip. We believe that the present work will facilitate the development of methods for the on-site diagnosis of diseases whose detection is based on antigen-antibody recognition, and improve the efficiencies of certain experiments that need to identify tens to hundreds of protein-protein interactions with relatively low cost in the future.