High-Throughput Mass Spectrometry Complements Protein Engineering

Protein research is critical in fundamental biochemistry, drug development, and biocatalyst applications. High-throughput discovery, characterization, and engineering of proteins are highly desirable, considering the overwhelming availability of genomic information that awaits functional annotation, and the enormous structure-function space that can be explored using protein engineering strategies. While one can now create a large number of engineered proteins using a variety of microbial expression systems, the ability to chemically characterize them in a high throughput manner has lagged behind. Capable of label-free analysis that utilizes native substrates and ligands, mass spectrometry (MS) is well suited for high-throughput assays to characterize complex mixtures of engineered proteins. Here, we review recent advances that improve the throughput and information content of MS-based protein assays and describe notable applications that highlight these capabilities.