Protein micropatterns printed on glass: Novel tools for protein‐ligand binding assays in live cells

Abstract

Micrometer‐sized patterns of proteins on glass or silica surfaces are in widespread use as protein arrays for probing with ligands or recombinant proteins. More recently, they have been used to capture the surface proteins of mammalian cells seeded onto them, and to arrange these surface proteins into pattern structures. Binding of small molecule ligands or of other proteins, transmembrane or intracellular, to these captured surface proteins can then be quantified. However, reproducible production of protein micropatterns on surfaces can be technically difficult. In this review, we outline the wide potential and the current practical uses of printed protein micropatterns in a historical overview, and we detail some potential pitfalls and difficulties from our own experience, as well as ways to circumvent them.

A two-hybrid antibody micropattern assay reveals specific in cis interactions of MHC I heavy chains at the cell surface

“…Silicon master molds were prepared by semiconductor photolithography as described previously. See Dirscherl et al (2017 ) for details.…”

“…Recently, antibody-based capture assays on solid supports have been described that can be used in bait-prey experiments in live cells ( Löchte et al, 2014 ; Schwarzenbacher et al, 2008 ; Weghuber et al, 2010 ). We now demonstrate an expansion of this concept to characterize location- and conformation-specific protein-protein interactions in a novel two-hybrid assay read out by fluorescence microscopy by using microprinted antibody patterns for the capture of bait proteins to spatially arrange them in the plasma membrane of live cells ( Dirscherl and Springer, 2017 ) and to investigate their interaction with green fluorescent protein (GFP)-tagged prey proteins. The assay is universally applicable for the investigation of protein-protein interactions.…”

Patterning of protein‐based materials

“…Micropatterned PDMS stamps represent a simple approach to transfer a protein of interest onto surfaces. [ 81–84 ] Recently, the simple soft lithography technique was used for creation of protein repellent/adhesive interfaces employing a marine mussels' adhesive mechanism. The marine mussels' tight binding to various substrates in aqueous environment relies on repeated 3,4‐dihydroxy‐l‐phenylalanine (DOPA) residues in mussel adhesive proteins, and DOPA patches provide a versatile tool for surface modifications.…”

Stable anchoring of bacteria-based protein nanoparticles for surface enhanced cell guidance

“…[6][7][8] Other approaches such as self-assembly and soft-lithography enabled versatile chemical functionalizations and immobilizations of biological materials on surfaces. [9][10][11][12][13] Among the techniques used to fabricate these soft materials, microcontact printing (mCP) stands as a cost-effective and fast soft-lithography technique to control surface chemistry at the microscale. 14 mCP allows the use of a broad range of materials to coat surfaces such as proteins, 15,16 nucleic acids, 17 genetically modified viruses, 18 and electroactive ligands with dynamic surfaces.…”