Main description:

This detailed volume presents a set of protocols useful for researchers in the field of recombinant immunoglobulin and alternative scaffold engineering, aptamer development, and generation of molecularly imprinted polymers (MIPs). Part I includes methods that deal with amino-acid based synthetic antibodies. Brief protocols about the generation of antibody libraries are detailed, as well as techniques for antibody selection, characterization, and validation. This section is completed by a brief description of a bioinformatics platform that supports antibody engineering during research and development. Part II contains basic procedures about the selection and characterization of aptamer molecules, and Part III describes fundamental processes of MIP generation and application. Written for the highly successful Methods in Molecular Biology series, 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.
Authoritative and practical, Synthetic Antibodies: Methods and Protocols is an ideal guide for scientists seeking to propel the vital study of antibody research.

Contents:

Part I: Amino Acid-Based Synthetic Antibodies

1. Antibody Mimetics, Peptides, and Peptidomimetics

Xiaoying Zhang and Thirumalai Diraviyam

2. Construction of a scFv Library with Synthetic, Non-Combinatorial CDR Diversity

Xuelian Bai and Hyunbo Shim

3. Enzymatic Assembly for scFv Library Construction

Mieko Kato and Yoshiro Hanyu

4. Directed Evolution of Protein Thermal Stability Using Yeast Surface Display

Michael W. Traxlmayr and Eric V. Shusta

5. Whole Cell Panning with Phage Display

Yvonne Stark, Sophie Venet, and Annika Schmid

6. Generating Conformation and Complex-Specific Synthetic Antibodies

Marcin Paduch and Anthony A. Kossiakoff

7. High-Throughput IgG Conversion of Phage Displayed Fab Antibody Fragments by AmplYFast

Andrea Sterner and Carolin Zehetmeier

8. Utilization of Selenocysteine for Site-Specific Antibody Conjugation

Xiuling Li and Christoph Rader

9. Solubility Characterization and Imaging of Intrabodies Using GFP-Fusions

Emilie Renaud, Pierre Martineau, and Laurence Guglielmi

10. Antibody Validation by Immunoprecipitation Followed by Mass Spectrometry Analysis

Helena Persson, Charlotta Preger, Edyta Marcon, Johan Lengqvist, and Susanne Graslund

11. Novel HPLC-Based Screening Method to Assess Developability of Antibody-Like Molecules

Neeraj Kohli and Melissa L. Geddie

12. Glycosylation Profiling of / T Cell Receptor Constant Domains Expressed in Mammalian Cells

Kai Zhang, Stephen J. Demarest, Xiufeng Wu, and Jonathan R. Fitchett

13. A Proximity-Based Assay for Identification of Ligand and Membrane Protein Interaction in Living Cells

Hongkai Zhang and Richard A. Lerner

14. A Biotin Ligase-Based Assay for the Quantification of the Cytosolic Delivery of Therapeutic Proteins

Wouter P.R. Verdurmen, Marigona Mazlami, and Andreas Pluckthun

15. Data-Driven Antibody Engineering Using Genedata Biologics (TM)

Maria Wendt and Guido Cappuccilli

Part II: Nucleotide-Based Synthetic Antibodies: Aptamers

16. Selection of Aptamers Against Whole Living Cells: From Cell-SELEX to Identification of Biomarkers

Nam Nguyen Quang, Anna Miodek, Agnes Cibiel, and Frederic Duconge

17. Rapid Selection of RNA Aptamers that Activate Fluorescence of Small Molecules

Grigory S. Filonov

18. An Enzyme-Linked Aptamer Sorbent Assay to Evaluate Aptamer Binding

Matthew D. Moore, Blanca I. Escudero, and Lee-Ann Jaykus

19. Incorporating Aptamers in the Multiple Analyte Profiling Assays (xMAP): Detection of C-Reactive Protein

Elyse D. Bernard, Kathy C. Nguyen, Maria C. DeRosa, Azam F. Tayabali, and Rocio Aranda-Rodriguez

Part III: Moleculary Imprinted Polymers

20. Transferring the Selectivity of a Natural Antibody into a Molecularly Imprinted Polymer

Romana Schirhagl

21. Preparation of Molecularly Imprinted Microspheres by Precipitation Polymerization

Tibor Renkecz and Viola Horvath

22. Generation of Janus Molecularly Imprinted Polymer Particles

Xiantao Shen and Chuixiu Huang

23. Surface Engineering of Nanoparticles to Create Synthetic Antibodies

Linda Chio, Darwin Yang, and Markita Landry

24. H5N1 Virus Plastic Antibody Based on Molecularly Imprinted Polymers

Chak Sangma, Peter A. Lieberzeit, and Wannisa Sukjee

25. Replacement of Antibodies in Pseudo-ELISAs: Molecularly Imprinted Nanoparticles for Vancomycin Detection

Francesco Canfarotta, Katarzyna Smolinska-Kempisty, and Sergey Piletsky

26. Cell and Tissue Imaging with Molecularly Imprinted Polymers

Maria Panagiotopoulou, Stephanie Kunath, Karsten Haupt, and Bernadette Tse Sum Bui