Protein arraying by cell-free expression and diffusion across a fluid-filled gap

Abstract

An improved system for cell-free expression of protein arrays based on DNA arrays is presented. Our technology uses an array of DNA constructs for cell-free expression, which acts as a template instructing the generation of the corresponding protein array. Proteins are expressed locally from these templates by a cell-free transcription and translation system, and are immobilised on a separate capture surface overlayed on the DNA array. By simplifying the setup to allow protein diffusion between the slides across a gap filled with the cell-free system, we have markedly improved the evenness of the resulting protein microarrays.