Abstract
•Prokaryotic expression platforms have been implemented in high-throughput structural biology environments.•Significant progress has been made towards realization of high-throughput eukaryotic expression platforms.•Lentiviral-driven and inducible systems are addressing challenging mammalian protein targets.•New methods are addressing heterogeneous glycosylation of secreted and cell surface molecules.•High-throughput technologies are having impact beyond traditional structural genomics communities.
Despite a multitude of recent technical breakthroughs speeding high-resolution structural analysis of biological macromolecules, production of sufficient quantities of well-behaved, active protein continues to represent the rate-limiting step in many structure determination efforts. These challenges are only amplified when considered in the context of ongoing structural genomics efforts, which are now contending with multi-domain eukaryotic proteins, secreted proteins, and ever-larger macromolecular assemblies. Exciting new developments in eukaryotic expression platforms, including insect and mammalian-based systems, promise enhanced opportunities for structural approaches to some of the most important biological problems. Development and implementation of automated eukaryotic expression techniques promises to significantly improve production of materials for structural, functional, and biomedical research applications.