Soft lithography-based biomolecule patterning techniques and their applications in subcellular protein interaction analysis

Soft lithography-based contact printing techniques have evolved into versatile methods for creating micro- and nanoscale features of biomolecules on solid substrates. In this review we present the advances in soft lithography for biomolecule deposition and its applications in subcellular protein-protein interaction (PPI) analysis. We discuss various soft lithography techniques, including micro-contact printing (μCP), nano-contact printing (nCP), capillary nanostamping, and polymer-pen-lithography (PPL) and focus on their application in biomolecule patterning on diverse substrates. We then address related challenges and advancements, including substrate selection, surface activation methods, and stamp development. The specific advantages, limitations, and potential solutions for printing various inks and biomolecules are highlighted. Furthermore, recent advances in soft lithography-based biomolecule patterning for subcellular protein interaction analysis are emphasized, demonstrating the importance of these techniques for incorporating complex cellular events into PPI readout modalities and established protein deposition strategies. Finally, an overview of future technologies and enhanced applications is provided.