Cell-Free Protein Synthesis for Protein Microarrays
Gregory A. Michaud, Michael Salcius, Rebecca Martone, Diane Buhr, John C. Duarte, Jennifer E. McCague, Xiangdong Liu, Michael Samuels, Christine Stalder, James Ball, Alex Tikhonov, Shiranthi Keppetipola, Wieslaw Kudlicki, James Meegan and Barry I. Schweitzer
Protein microarray technology has empowered investigators with a tool that offers the potential to study the function of thousands of proteins in a single experiment. The approach typically involves a process by which proteins are tethered to either a modified microscope slide or to the bottom of a well in a multi-well plate. Numerous applications have been developed that measure interactions, both covalent and noncovalent, of a probe with target proteins on the array. Almost every type of interaction observed in biomolecular signaling pathways has been reconstituted on protein arrays, including interactions with small molecules, lipids, proteins, DNA, RNA, carbohydrate and enzymes.1,2 One of the most common questions related to adoption of the technology as a complementary platform for research and development is, “How will protein microarray technology bring value to my research?” One answer to this question is that protein microarray technology can greatly accelerate the speed at which new information can be generated. Standard approaches to investigating protein function can take months to years to address the function of just one protein. In contrast, a standard protein microarray experiment can be performed in as little as 5 hours. Because the technology enables such rapid screening, the cost savings in materials and labor can be significant. Secondly, screens on protein microarrays can be comprehensive in that an entire proteome can be profiled in a single experiment which is generally not achievable by alternate proteomic technologies. Below, we highlight some recent examples of how protein microarray technology has been applied to further understand the biology of complex systems. In this discussion, we will focus on the application of cell-free protein synthesis and how it has been applied in protein microarray experiments.