Recombinant Protein Expression: Eukaryotic hosts

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.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.

Recombinant Protein Expression, Part B, Volume 660 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more.

Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae.

Preface

1. Expression of recombinant multi-protein complexes in Saccharomyces cerevisiae

2. Recombinant production of membrane proteins in yeast

3. Expression of proteins in Pichia pastoris

4. Hybrid-architectured promoter design to engineer expression in yeast

5. Hybrid-architectured promoter design to deregulate expression in yeast

Baculovirus-insect cell systems

6. The MultiBac BEVS: Basics, Applications, Performance and Recent Developments

7. Applications of Golden Gate Cloning to protein production using the baculovirus expression vector system

8. A fast-track protocol for protein expression using the BEV system

Plant hosts

9. Transient expression of recombinant proteins in plants

10. Transient expression of recombinant proteins in plants using potato virus X based vectors

11. Design and expression of a bispecific antibody against dengue and chikungunya virus in plants

12. Manufacturing plant-made monoclonal antibodies for research or therapeutic applications

Mammalian cell lines as biomanufacturing platforms

13. Creation of monoclonal antibody expressing CHO cell lines grown with sodium butyrate and characterization of resulting antibody glycosylation

14. Method to transfer Chinese hamster ovary (CHO) batch shake flask experiments to large-scale, computer-controlled fed-batch bioreactors

15. Inducible protein expression in piggyBac transposase mediated stable HEK293 cell pools

16. Exploring the design space of AAV transient-transfection in suspension cell lines