In this paper we present an experimental protocol for protein immobilization on polydimethylsiloxane (PDMS) polymer surfaces and the subsequent application of a chromatographic PDMS microfluidic chip to measure protein-protein interactions. The PDMS surface modification steps are quantitatively and qualitatively experimentally analyzed using an array of techniques (water contact angle measurement, fluorescence spectroscopy and X-ray photoelectron spectroscopy). The protocol involves PDMS acidic surface activation using a potassium disulfite/potassium peroxidisulfate/acrylic acid mixture, followed by amination with 3-aminopropyl diethoxymethylsilane, followed by glutaraldehyde grafting and subsequent covalent protein binding. The applicability of such a miniaturized PDMS-based microfluidic system has been exemplified by measuring protein-protein interactions in a fast and accurate fashion for three model proteins, namely: hen egg white lysozyme, bovine ribonuclease-A and α-chymotrypsinogen. The protein interaction results align well with existing literature data using different materials and techniques. As the fabrication process for PDMS-based microstructures is relatively cheap, quick and requires limited lab expertise/access to specialized equipments, we consider that the implementation of such a flexible, easy to fabricate, PDMS-based microfluidic system for estimating protein interactions an important step toward quickly mapping protein phase behavior and measuring protein (self/cross) interactions in complex biological systems.
Deshpande, K.S., S. Kuddannaya, J. Staginus, P.C. Thune, L.C.P.M. de Smet, J.H. ter Horst, L.A.M. van der Wielen, M. Ottens
Biochemical Engineering Journal