High-Throughput Protein Production and Purification: Methods and Protocols

ISBN-13: 9781493996230 / Angielski / Twarda / 2019 / 538 str.

Renaud Vincentelli

Spis treści

Part 1 General Procedures

1)      HTP cloning

2)      HTP generation of truncation and mutation libraries

3)      HTP protein production in Escherichia coli

4)      HTP Production of Protein Complexes by Co-expression in Escherichia coli

5)      HTP protein complex reconstitution in Escherichia coli

6)      HTP protein production in alternative bacterial system

7)      HTP protein production in Yeast

8)      HTP protein production and protein complexes in Baculovirus

9)      HTP protein production and protein complexes in Mammalian cells

10)  HTP protein production in plants

11)  HTP protein expression in bacterial Cell Free system

12)  HTP protein expression in eukaryotic Cell Free system

13)  HTP refolding of proteins

14)  HTP production of binders

15)  HTP production of antibodies

16)  HTP Modification of strains for improved protein expression (CHO, HEK, E. coli)

Part 2 Case studies

17)  HTP production of membrane proteins

18)  HTP membrane protein expression in bacterial Cell Free system

19)  HTP production of kinases

20)  HTP production of small cysteine-rich proteins (VENOMICS)

21)  HTP production of transcription factors

22)  HTP Protein production for drug discovery

Part 3 HTP micro-characterizations of proteins

23)  Analytical SEC- MALLS

24)  Differential Scanning Fluorimetry

25)  Native mass spectrometry

26)  Protein-protein interaction (Pull-down/Hold-up)

27)  Protein-DNA (SELEX)

28)  Analytical Electron microscopy

Blurb

This book compiles key protocols instrumental to the study of high-throughput protein production and purification which have been refined and simplified over the years and are now ready to be transferred to any laboratory. Beginning with a section covering general procedures for high-throughput protein production, the volume continues with high-throughput protocols adapted to the production of specific protein families, as well as an extensive section on protocols combining high-throughput protein production and their micro-characterization. Written for the highly successful Methods in Molecular Biology series, chapters in this book include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and practical, High-Throughput Protein Production and Purification: Methods and Protocols serves biochemists ranging from engineers, PhD students and post-doctoral fellows, to the heads of protein expression facilities and researchers, in pursuing this vital area of study.