Printing Proteins as Microarrays for High-Throughput Function Determination

TLDR

Efforts are underway to build defined gene sets for high‑throughput recombinant protein expression and purification. The study aims to develop miniaturized assays that use very low sample volumes to enable rapid, simultaneous processing of thousands of proteins for functional studies. A high‑precision robotic system spots proteins onto chemically derivatized glass slides at extremely high spatial densities. Proteins covalently bound to the slides retained specific binding activity, enabling applications in protein‑protein interaction screening, kinase substrate identification, and small‑molecule target discovery.

Abstract

Systematic efforts are currently under way to construct defined sets of cloned genes for high-throughput expression and purification of recombinant proteins. To facilitate subsequent studies of protein function, we have developed miniaturized assays that accommodate extremely low sample volumes and enable the rapid, simultaneous processing of thousands of proteins. A high-precision robot designed to manufacture complementary DNA microarrays was used to spot proteins onto chemically derivatized glass slides at extremely high spatial densities. The proteins attached covalently to the slide surface yet retained their ability to interact specifically with other proteins, or with small molecules, in solution. Three applications for protein microarrays were demonstrated: screening for protein-protein interactions, identifying the substrates of protein kinases, and identifying the protein targets of small molecules.