Reagent-Free Covalent Immobilization of Biomolecules in a Microfluidic Organ-On-A-Chip

Microfluidic systems have become integral for lab-on-a-chip and organ-on-a-chip applications across numerous disciplines. These systems, typically fabricated using polydimethylsiloxane (PDMS) chips on glass substrates, lack the bioactivity required for such applications. To overcome this, biomolecules are immobilized using either oxygen (O2) plasma treatment or chemical reagents like amino silanes. However, O2 plasma treatments are unstable and cannot covalently immobilize biomolecules, while wet-chemistry approaches are toxic, time-consuming, and expensive. A novel microfluidic platform that combines two plasma surface treatments: Plasma-activated coating (PAC) and atmospheric pressure plasma jet (APPJ), to enable reagent-free covalent immobilization of biomolecules is described here. These surface treatments, unlike O2 plasma, covalently immobilized fibronectin on PDMS and glass, and significantly improved endothelial cell attachment and proliferation. By combining PAC and APPJ, a hybrid microfluidic platform with equivalent bond strength to standard O2 plasma devices, but with significantly enhanced endothelial cell growth in and artery-on-a-chip model, is developed. This platform is also amenable to high-shear applications such as coronary shear, with endothelial cells aligning with flow, as seen in human arteries. By providing reagent-free covalent immobilization of biomolecules within a microfluidic system, this technology has the potential to radically improve organ-on-a-chip development as well as lab-on-a-chip systems, point-of-care diagnostics, and sensors. By combining the novel plasma treatment techniques PAC and APPJ on PDMS and glass respectively, a microfluidic platform is developed, that for the first time enables reagent-free covalent immobilization of biomolecules. Using this platform, improved endothelial cell growth is demonstrated relative to standard oxygen plasma treatments under both low and high-shear conditions for multiple days. image