Recombinant Protein Expression: Prokaryotic hosts and cell-free systems

William Brad O'Dell is a PhD biologist experienced in in Molecular Biology, Protein Expression, Protein Purification, Crystallography, Lab-Scale Yeast Fermentation, Flow Cytometry and Small Angle Scattering of Molecules and Materials. Zvi Kelman is the Director of the Biomolecular Labeling Laboratory (BL2), National Institute of Standards and Technology (NIST). He is also an Adjunct Professor in the Department of Cell Biology and Molecular Genetics at the University of Maryland, College Park and affiliated with the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine. Zvi earned a B.Sc. in Agriculture from the Hebrew University in Jerusalem and a M.Sc. in Cell Biology from the Weizmann Institute of Science. After receiving a Ph.D. in Molecular Biology from Cornell University, he was a Helen Hay Whitney Foundation Post-Doctoral Fellow at Johns Hopkins University and Memorial Sloan-Kettering Cancer Center. Upon completion of his postgraduate training, he became a Life Technologies Professor at the Center for Advanced Research in Biotechnology (CARB), University of Maryland Biotechnology Institute (UMBI). Zvi moved to the University of Maryland, College Park in 2010 as a Professor in the Department of Cell Biology and Molecular Genetics. In 2011 he was recruited to NIST to direct the Biomolecular Labeling Laboratory.

• PrefaceBacterial hosts1. Starting a new recombinant protein production project in Escherichia coli2. From the notebook to recombinant protein production in Escherichia coli: design of expression vectors and gene cloning3. Use of tandem affinity–buffer exchange chromatography online with native mass spectrometry for optimizing overexpression and purification of recombinant proteins4. Purification, reconstitution, and mass analysis of archaeal RNase P, a multi-subunit ribonucleoprotein enzyme5. Production of antibodies in SHuffle E. coli strains6. Improved folding of recombinant protein via co-expression of exogenous chaperones7. Fusing an insoluble protein to GroEL apical domain enhances soluble expression in Escherichia coli8. Method for high-efficiency fed-batch cultures of recombinant Escherichia coli9. Fed-batch production of deuterated protein in Escherichia coli for neutron scattering experimentationArchaeal hosts10. Thermococcus kodakarensis provides a versatile hyperthermophilic archaeal platform for protein expression11. Recombinant protein expression in Sulfolobus islandicus12. High-level synthesis and secretion of laccase, a metalloenzyme biocatalyst, by the halophilic archaeon Haloferax volcanii13. Tandem affinity purification of 20S proteasomes and other multisubunit complexes in Haloferax volcanii14. Purification and characterization of ribonucleoprotein effector complexes of Sulfolobus islandicus CRISPR-Cas systemsCell-free systems15. Guidelines for nucleic acid template design for optimal cell-free protein synthesis using an E. coli reconstituted system or a lysate-based system16. Cell-free protein synthesis of CRISPR ribonucleoproteins (RNP)17. Leishmania tarentolae cell-free based approach for rapid antibody: antigen interaction analysis18. Cell-free protein synthesis using Chinese hamster ovary cells