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Ribosome Display and Related Technologies By Julie A. Douthwaite

Ribosome Display and Related Technologies by Julie A. Douthwaite

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Ribosome Display and Related Technologies Summary

Ribosome Display and Related Technologies: Methods and Protocols by Julie A. Douthwaite

Display technologies have become a very powerful way of generating therapeutic lead molecules and specific reagents for increasing our understanding of biology; however, despite being first described shortly after phage display, the use of ribosome display and related methods have been much less widespread. Since this is in part due to the complexity of the methods, Ribosome Display and Related Technologies: Methods and Protocols seeks to extend their use by collecting expert contributions describing these detailed protocols. The protocols described range from well-established methods that have been used for a decade to generate high affinity antibodies, which are already in the clinic, to methods that are in their early stages of application such as display of peptides incorporating non-canonical amino acids. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters 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.

Invaluable and easy to use, Ribosome Display and Related Technologies: Methods and Protocols will be of great benefit to those with general molecular biology or protein engineering experience who wish to select peptides or proteins by display, those with phage display experience who would benefit from the application of ribosome display, as well as those with some ribosome display experience who would like to expand the range of applications to which they are applying the technology.

Table of Contents

Part I: Review

1. Ribosome Display: A Perspective

Andreas Pluckthun

Part II: Translation Extract Preparation

2. Preparation and Testing of E.coli S30 In Vitro Transcription Translation Extracts

James F. Zawada

Part III: Basic Ribosome Display and Related Selection Methods

3. Eukaryotic Ribosome Display Selection Using Rabbit Reticulocyte Lysate

Julie A. Douthwaite

4. Stabilized Ribosome Display for In Vitro Selection

Shuta Hara, Mingzhe Liu, Wei Wang, Muye Xu, Zha Li, and Yoshihiro Ito

5. Eukaryotic Ribosome Display with In Situ DNA Recovery

Mingyue He, Bryan M. Edwards, Damjana Kastelic, and Michael J. Taussig

6. mRNA Display Using Covalent Coupling of mRNA to Translated Proteins

Rong Wang, Steve W. Cotten, and Rihe Liu

7. SNAP Display: In Vitro Protein Evolution in Microdroplets

Miriam Kaltenbach and Florian Hollfelder

8. cDNA Display: Rapid Stabilization of mRNA Display

Shingo Ueno and Naoto Nemoto

Part IV: Applications of Ribosome Display Methods Using Natural Amino Acids

9. Optimization of Antibody Affinity by Ribosome Display Using Error-Prone or Site-Directed Mutagenesis

Leeanne Lewis and Chris Lloyd

10. Affinity Maturation of Phage Display Antibody Populations using Ribosome Display

Maria A. Groves and Adrian A. Nickson

11. Evolution of Protein Stability Using Ribosome Display

Andrew Buchanan

12. Selection of Lead Antibodies from Naive Ribosome Display Antibody Libraries

Peter Ravn

13. Evolution of Disulfide-Rich Peptide Aptamers Using cDNA Display

Yuki Mochizuki and Naoto Nemoto

14. Peptide Screening Using PURE Ribosome Display

Hiroyuki Ohashi, Takashi Kanamori, Eriko Osada, Bintang K. Akbar, and Takuya Ueda

15. Rapid Selection of High Affinity Binders Using Ribosome Display

Birgit Dreier and Andreas Pluckthun

16. mRNA Display-Based Selections Using Synthetic Peptide and Natural Protein Libraries

Steve W. Cotten, Jianwei Zou, Rong Wang, Bao-cheng Huang, and Rihe Liu

17. Identification of Candidate Vaccine Genes Using Ribosome Display

Liancheng Lei

18. Ribosome Display for the Selection of Sac7d Scaffolds

Barbara Mouratou, Ghislaine Behar, Lauranne Paillard-Laurance, Stephane Colinet, and Frederic Pecorari

Part V: Incorporation of Non-Natural Amino Acids for Selection by Ribosome Display and Related Methods

19. Charging of tRNAs Using Ribozymes and Selection of Cyclic Peptides Containing Thioethers

Patrick C. Reid, Yuki Goto, Takayuki Katoh, and Hiroaki Suga

20. Update on Pure Translation Display with Unnatural Amino Acid Incorporation

R. Edward Watts and Anthony C. Forster

21. In Vitro Selection of Unnatural Cyclic Peptide Libraries Via mRNA Display

Zhong Ma and Matthew C.T. Hartman

Part VI: Case Studies

22. Optimization of CAT-354, a Therapeutic Antibody Directed Against Interleukin-13, Using Ribosome Display

George Thom and Ralph Minter

23. Affinity Maturation and Functional Dissection of a Humanized Anti-RAGE Monoclonal Antibody by Ribosome Display

Simon E. Hufton

Additional information

Ribosome Display and Related Technologies: Methods and Protocols by Julie A. Douthwaite
Humana Press Inc.
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