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Dominic Esposito, Ph.D.
Director, Protein Expression Laboratory
Program Overview
The Protein Expression Laboratory generates DNA, cell line, and protein reagents for biomedical research from basic science to drug discovery. The team uses cutting-edge protein production technologies to generate proteins using bacteria, insect cells, or mammalian cells at scales from micrograms to grams. Quality control is a major emphasis of the laboratory, ensuring that all protein reagents pass a series of stringent tests before leaving the group. The laboratory also focuses on standardization and improvement to protein production technologies that are shared with the extramural research community.
Program Innovations
- The Protein Expression Laboratory has made substantial improvements to the production of vital COVID-19 serology reagents in support of NIH efforts.
- The laboratory has generated a variety of novel protein production technologies to both improve protein yields and quality, and reduce the time needed to generate protein reagents.
Program Focus
Proteins for structural biology applications
- Produce high-quality individual recombinant proteins and protein complexes for use in X-ray crystallography, nuclear magnetic resonance, and cryo-electron microscopy.
- Specialize in optimization of protein constructs using microscale expression and purification to quickly identify the optimal protein domains for production.
- For advanced structural biology work, proteins can readily be labelled with isotopes for nuclear magnetic resonance structural determination or heavy metals for X-ray crystallography optimization.
Proteins to support drug discovery assay development and deployment
- Generate proteins with a variety of domain designs and tagging strategies to support all types of biochemical and biophysical drug discovery assays.
- Maintain a collection of more than 100 different fusion protein tags ranging from various fluorescent proteins to antibody-based detection tags.
- Proteins have been used for assays including surface plasmon resonance, homogenous time-resolved fluorescence, fluorescence polarization, AlphaScreen, and isothermal calorimetry.
Creation of new technologies for enhanced protein production
- Develop new reagents to improve production of proteins from insect cells, including:
- Novel high-producing cell lines
- Modified DNA constructs for faster and more efficient baculovirus production
- Chaperones to assist with better recombinant protein folding
- Improve protein purification within the group, including the use of magnetic bead technologies to accelerate protein purification timelines and reduce cost, as well as new expression systems to provide additional options for challenging protein targets
- Use specialized cloning processes to permit multi-gene assembly assist in the production of large multiprotein complexes for structural biology and drug discovery