📖 Protein microarray synthesis and label-free interaction analysis. Design and evaluation of automated system.

Protein microarrays are essential tools for high-throughput proteome interaction and binding kinetics analysis. Recent technical developments have resulted in approaches for the generation of protein arrays by in situ cell-free expression of DNA array templates. We combined and enhanced these ideas by creating innovative microfluidic flow cells in standard microscope slide format and such improved system efficiency, operability, robustness and controllability. An automated setup suitable for both the in situ expression as well as the interaction analysis of protein microarrays by flow-injection was designed, built and evaluated. The reflecto-interferometric principle was selected to enable high-throughput label-free imaging detection. Both, the protein microarray expression and the microarray interaction analysis could be performed sequentially in the same microfluidic flow cell - the created proteins remain in their fluidic environment and never get in contact with air. Various antibody-antigene and rabbit-sera immunization assays were proving this system to be an efficient tool for interaction screenings.