Recombinant Protein Expression, Part A, Volume 659 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.
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Table of Contents
PrefaceBacterial hosts
1. Starting a new recombinant protein production project in Escherichia coli
2. From the notebook to recombinant protein production in Escherichia coli: design of expression vectors and gene cloning
3. Use of tandem affinity-buffer exchange chromatography online with native mass spectrometry for optimizing overexpression and purification of recombinant proteins
4. Purification, reconstitution, and mass analysis of archaeal RNase P, a multi-subunit ribonucleoprotein enzyme
5. Production of antibodies in SHuffle E. coli strains
6. Improved folding of recombinant protein via co-expression of exogenous chaperones
7. Fusing an insoluble protein to GroEL apical domain enhances soluble expression in Escherichia coli
8. Method for high-efficiency fed-batch cultures of recombinant Escherichia coli
9. Fed-batch production of deuterated protein in Escherichia coli for neutron scattering experimentation
Archaeal hosts
10. Thermococcus kodakarensis provides a versatile hyperthermophilic archaeal platform for protein expression
11. Recombinant protein expression in Sulfolobus islandicus
12. High-level synthesis and secretion of laccase, a metalloenzyme biocatalyst, by the halophilic archaeon Haloferax volcanii
13. Tandem affinity purification of 20S proteasomes and other multisubunit complexes in Haloferax volcanii
14. Purification and characterization of ribonucleoprotein effector complexes of Sulfolobus islandicus CRISPR-Cas systems
Cell-free systems
15. Guidelines for nucleic acid template design for optimal cell-free protein synthesis using an E. coli reconstituted system or a lysate-based system
16. Cell-free protein synthesis of CRISPR ribonucleoproteins (RNP)
17. Leishmania tarentolae cell-free based approach for rapid antibody: antigen interaction analysis
18. Cell-free protein synthesis using Chinese hamster ovary cells