Genotype Phenotype Coupling Methods and Protocols

This volume aims at providing state-of-the-art protocols detailing ribosome display, cDNA display, phage display, yeast surface display, and mammalian display. Chapters guide readers through methods and protocols on in vitro methods over prokaryotic display systems, lower eukaryotes, and mammalian cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of all necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Genotype Phenotype Coupling aims to provide an overview of current technologies in this exciting and continuously evolving field. 

1. Screening, Post-SELEX Optimization and Application of DNA Aptamers Specific for Tobramycin

Nandi Zhou, Rongfeng Cai, and Xuyan Han

2. Affitins: Ribosome Display for Selection of Aho7c-based Affinity Proteins

Valentina Kalichuk, Stanimir Kambarev, Ghislaine Béhar, Benjamin Chalopin, Axelle Renodon-Cornière, Barbara Mouratou, and Frédéric Pecorari

3. cDNA Display: A Stable and Simple Genotype–phenotype Coupling Using a Cell-free Translation System

Hidenao Arai, Shigefumi Kumachi, and Naoto Nemoto

4. cDNA Display of Disulfide-containing Peptide Library and in vitro Evolution

Tai Kubo and Mohammed Naimuddin

5. Rapid Antigen and Antibody-like Molecule Discovery by Staphylococcal Surface Display

Marco Cavallari

6.  Restriction-free Construction of a Phage-presented Very Short Macrocyclic Peptide Library

Valentin Jakob, Saskia Helmsing, Michael Hust, and Martin Empting

7. In Vitro Maturation of a Humanized Shark VNAR Domain to Improve Its Biophysical Properties

8. Antibody Phage Display: Antibody Selection in Solution Using Biotinylated Antigens 

Esther V. Wenzel, Kristian D. R. Roth, Giulio Russo, Viola Fühner, Saskia Helmsing, André Frenzel, and Michael Hust

9. Assessing Antibody Specificity in Human Serum Using Deep Sequence-coupled Biopanning

Kathryn M. Frietze, Susan B. Core, Alexandria Linville, Bryce Chackerian, and David S. Peabody

10. Isolation of Antigen-specific VHH Single Domain Antibodies by Combining Animal Immunization with Yeast Surface DisplayLukas Roth, Simon Krah, Janina Klemm, Ralf Guenther, Lars Toleikis, Michael Busch, Stefan Becker, and Stefan Zielonka 

11. Selection and Characterization of Anti-idiotypic Shark Antibody Domains

Doreen Könning, Stefan Zielonka, Anna Kaempffe, Sebastian Jäger, Harald Kolmar, and Christian Schröter

12. Simplifying the Detection of Surface Presentation Levels in Yeast Surface Display by Intracellular tGFP Expression

Steffen C. Hinz, Adrian Elter, Julius Grzeschik, Jan Habermann, Bastian Becker, and Harald Kolmar

13. Methods for Construction of Yeast Display Libraries of Four-Domain T-cell Receptors

Flávio Sádio, Gerhard Stadlmayr, Katja Eibensteiner, Katharina Stadlbauer, Florian Rüker, and Gordana Wozniak-Knopp

14.  Isolation of Tailor-made Antibody Fragments from Yeast-displayed B-Cell Receptor Repertoires by Multiparameter Fluorescence-Activated Cell Sorting

Anna Kaempffe, Sebastian Jäger, Doreen Könning, Harald Kolmar, and Christian Schröter

15. Isolation of Anti-Hapten Antibodies by Fluorescence-Activated Cell Sorting of Yeast-Displayed B-Cell Receptor Gene Repertoires

Sebastian Jäger, Simon Krah, Doreen Könning, Janis Rosskopf, Stephan Dickgiesser, Nicolas Rasche, Harald Kolmar, Stefan Hecht, and Christian Schröter

16. Rapid Generation of Chicken Immune Libraries for Yeast Surface Display

Jan P. Bogen, Julius Grzeschik, Simon Krah, Stefan Zielonka, and Harald Kolmar

17. Ligand Engineering via Yeast Surface Display and Adherent Cell Panning

Lawrence A. Stern, Patrick S. Lown, and Benjamin J. Hackel

18. Simultaneous Soluble Secretion and Surface Display of Proteins in Saccharomyces Cerevisiae Using Inefficient Ribosomal Skipping

Carlos A. Cruz-Teran, Kaitlyn Bacon, and Balaji M. Rao

19. Chemical Modification of the Yeast Cell Surface Allows the Switch between Display and Soluble Secretion of Full-length Antibodies

Simon Krah, Ralf Günther, Stefan Becker, Stefan Zielonka, and Laura Rhiel

20. Advanced Establishment of Stable Recombinant Human Suspension Cell Lines Using Geno-Phenotype Coupling Transposon Vectors

Karen Berg, Vanessa Nicole Schäfer, Natalie Bartnickiand,  and Jörn Stitz

Zachary R. Crook, Gregory P. Sevilla, Andrew J. Mhyre, and James M. Olson

22. Engineering Antibodies on the Surface of CHO Cells

Annalee W. Nguyen, Kevin Le, and Jennifer A. Maynard

Juliet Rashidian and Joshua Lloyd