Combinatorial Peptide Library Protocols / Edition 1

by Shmuel Cabilly

Overview

During the course of evolution, an imbalance was created between the rate of vertebrate genetic adaptation and that of the lower forms of living organisms, such as bacteria and viruses. This imbalance has given the latter the advantage of generating, relatively quickly, molecules with unexpected structures and features that carry a threat to vertebrates. To compensate for their weakness, vertebrates have accelerated their own evolutionary processes, not at the level of whole organism, but in specialized cells containing the genes that code for antibody molecules or for T-cell receptors. That is, when an immediate requirement for molecules capable of specific interactions arose, nature has preferred to speed up the mode of Darwinian evolution in pref- ence to any other approach (such as the use of X-ray diffraction studies and computergraphic analysis). Recently, Darwinian rules have been adapted for test tube research, and the concept of selecting molecules having particular characteristics from r- dom pools has been realized in the form of various chemical and biological combinatorial libraries. While working with these libraries, we noticed the interesting fact that when combinatorial libraries of oligopeptides were allowed to interact with different selector proteins, only the actual binding sites of these proteins showed binding properties, whereas the rest of the p- tein surface seemed "inert. " This seemingly common feature of protein- having no extra potential binding sites—was probably selected during evolution in order to minimize nonspecific interactions with the surrounding milieu.

Product Details

ISBN-13:	9781617370229
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/10/2010
Series:	Methods in Molecular Biology , #87
Edition description:	Softcover reprint of hardcover 1st ed. 1998
Pages:	313
Product dimensions:	5.98(w) x 9.02(h) x 0.35(d)

About the Author

Cabilly, Shmuel (Weizmann Institute of Science)

Table of Contents

Synthesis of a One-Bead One-Compound Combinatorial Peptide Library.- Enzyme-Linked Calorimetric Screening of a One-Bead One-Compound Combinatorial Library.- Synthesis and Screening of Positional Scanning Combinatorial Libraries.- Synthesis and Screening of Peptide Libraries on Continuous Cellulose Membrane Supports.- Peralkylation.- to Combinatorial Solid-Phase Assays for Enzyme Activity and Inhibition.- Preparation of Biocompatible Resins for Library Syntheses.- Intramolecular Fluorescence-Quenched Substrate Libraries.- The Solid-Phase Enzyme Inhibitor Library Assay.- Determination of Peptide Substrate Motifs for Protein Kinases Using a “One-Bead One-Compound” Combinatorial Library Approach.- The Use of Peptide Library for the Determination of Kinase Peptide Substrates.- Analysis of Protein Kinase Substrate Specificity by the Use of Peptide Libraries on Cellulose Paper (SPOT-Method).- Generation of Multiuse Peptide Libraries for Functional Screenings.- Functional Screening of Multiuse Peptide Libraries Using Melanophore Bioassay.- The Basic Structure of Filamentous Phage and Its Use in the Display of Combinatorial Peptide Libraries.- Construction and Use of a 20-mer Phage Display Epitope Library.- Construction of Disulfide-Constrained Random Peptide Libraries Displayed on Phage Coat Protein VIII.- Conformational Mimicry Through Random Constraints Plus Affinity Selection.- The Use of Combinatorial Libraries to Identify Ligands That Interact with Surface Receptors in Living Cells.- Screening Phage Display Peptide Libraries on Nitrocellulose Membranes.- Identification of Disease-Specific Epitopes.- Identification of Peptide Ligands for the Antigen Binding Receptor Expressed on Human B-Cell Lymphomas.- Major Hisompatibility Complex Allele-Specific Peptide Librariesand Identification of T-Cell Mimotopes.- Phage Display of Peptide Libraries on Protein Scaffolds.- Displaying Libraries of Conformationally Constrained Peptides on the Surface of Escherichia coli as Flagellin Fusions.- Combinatorial Peptide Library as an Immunogen.

Editorial Reviews

3 Stars from Doody

This is a timely book that covers many of the basic principles and methods of construction and use of peptide libraries. The initial chapters describe methods for the chemical synthesis of peptide libraries, and their use in screening for ligands. The intermediate chapters deal with biological libraries that are displayed using phage or bacteria. The final chapters are devoted to some practical applications of combinatorial library technology. This is a fast-growing area, which has been developing quickly since it was introduced in 1990. Many of the chapters delve into new areas such as "one-bead one-compound peptide libraries," "split synthesis," and the "SPOT method." This book is written by many recognized and respected scientists in the field. It is aimed primarily at those already familiar with the methodologies behind combinatorial peptide library synthesis and their practical applications. It is a nice handbook for those planning to use the covered techniques in the lab. Each of the chapters contains appropriate illustrations or photos where needed to express an idea. The references are both current and pertinent. This book will be very useful to both those who are just beginning working in this area as well as those already working in it but looking to expand with new approaches. However, some important methods such as plasmid display are not presented at all. Overall though, I would recommend it for a library or individual interested in combinatorial techniques.

Annette Gilchrist

A collection of new techniques for the construction and use of peptide libraries, with much of the material authored by pioneers in the methods. Offers step-by-step instructions for protocols for the chemical synthesis of peptide libraries, for constructing peptide libraries displayed on the surface of filamentous phage or bacteria, and for rapid screening of these libraries for molecules with biospecific properties. Additional methods permit identification of enzyme substrates, investigation of recognition spectra of various binding proteins, epitope mapping, and identification of mimotopes. Includes b&w diagrams. For molecular biologists, chemists, pharmacologists, bacteriologists, virologists, and immunologists. Annotation c. by Book News, Inc., Portland, Or.

Booknews

Reviewer: Annette Gilchrist, PhD (Northwestern University )
Description: This is a timely book that covers many of the basic principles and methods of construction and use of peptide libraries.
Purpose: The initial chapters describe methods for the chemical synthesis of peptide libraries, and their use in screening for ligands. The intermediate chapters deal with biological libraries that are displayed using phage or bacteria. The final chapters are devoted to some practical applications of combinatorial library technology. This is a fast-growing area, which has been developing quickly since it was introduced in 1990. Many of the chapters delve into new areas such as "one-bead one-compound peptide libraries," "split synthesis," and the "SPOT method."
Audience: This book is written by many recognized and respected scientists in the field. It is aimed primarily at those already familiar with the methodologies behind combinatorial peptide library synthesis and their practical applications. It is a nice handbook for those planning to use the covered techniques in the lab.
Features: Each of the chapters contains appropriate illustrations or photos where needed to express an idea. The references are both current and pertinent.
Assessment: This book will be very useful to both those who are just beginning working in this area as well as those already working in it but looking to expand with new approaches. However, some important methods such as plasmid display are not presented at all. Overall though, I would recommend it for a library or individual interested in combinatorial techniques.

Doody's Review Service