ADVANCES IN IMMUNOASSAY TECHNOLOGY docx

ADVANCES IN IMMUNOASSAY TECHNOLOGY Edited by Norman H. L. Chiu and Theodore K. Christopoulos Advances in Immunoassay Technology Edited by Norman H. L. Chiu and Theodore K. Christopoulos Published by InTech Janeza Trdine 9, 51000 Rijeka, Croatia Copyright © 2012 InTech All chapters are Open Access distributed under the Creative Commons Attribution 3.0 license, which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. After this work has been published by InTech, authors have the right to republish it, in whole or part, in any publication of which they are the author, and to make other personal use of the work. Any republication, referencing or personal use of the work must explicitly identify the original source. As for readers, this license allows users to download, copy and build upon published chapters even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. Notice Statements and opinions expressed in the chapters are these of the individual contributors and not necessarily those of the editors or publisher. No responsibility is accepted for the accuracy of information contained in the published chapters. The publisher assumes no responsibility for any damage or injury to persons or property arising out of the use of any materials, instructions, methods or ideas contained in the book. Publishing Process Manager Maja Bozicevic Technical Editor Teodora Smiljanic Cover Designer InTech Design Team First published March, 2012 Printed in Croatia A free online edition of this book is available at www.intechopen.com Additional hard copies can be obtained from orders@intechopen.com Advances in Immunoassay Technology, Edited by Norman H. L. Chiu and Theodore K. Christopoulos p. cm. ISBN 978-953-51-0440-7 Contents Preface IX Part 1 New Materials and Assay Interference 1 Chapter 1 Recombinant Antibodies and Non-Antibody Scaffolds for Immunoassays 3 Bhupal Ban and Diane A. Blake Chapter 2 Polyacrylonitrile Fiber as Matrix for Immunodiagnostics 23 Swati Jain, Sruti Chattopadhyay, Richa Jackeray, Zainul Abid and Harpal Singh Chapter 3 Interferences in Immunoassays 45 Johan Schiettecatte, Ellen Anckaert and Johan Smitz Part 2 Label-Free Technologies 63 Chapter 4 Fundamentals and Applications of Immunosensors 65 Carlos Moina and Gabriel Ybarra Chapter 5 Capabilities of Piezoelectric Immunosensors for Detecting Infections and for Early Clinical Diagnostics 81 Tatyana Ermolaeva and Elena Kalmykova Chapter 6 Label-Free Detection of Botulinum Neurotoxins Using a Surface Plasmon Resonance Biosensor 109 Hung Tran and Chun-Qiang Liu Chapter 7 Immunoassays Using Artificial Nanopores 125 Paolo Actis, Boaz Vilozny and Nader Pourmand Part 3 Multiplexing Technologies 141 Chapter 8 Multiplexed Immunoassays 143 Weiming Zheng and Lin He VI Contents Chapter 9 Multiplexed Bead Immunoassays: Advantages and Limitations in Pediatrics 165 Emma Burgos-Ramos, Gabriel Ángel Martos-Moreno, Jesús Argente, Vicente Barrios Preface Over the past decade, the development and applications of immunoassays have continued to grow exponentially. This book focuses on some of the latest advances in immunoassay technology, which include new materials and methods. The book contains nine invited chapters that are divided into three sections. In the first section, the basics for producing recombinant antibodies, the use of polyacrylonitrile fibre as a solid surface, and the nature of interference in immunoassays are summarized. The second section begins with a chapter on the basic concepts of different types of immunosensors, some of which allow label-free detection of specific analytes. This is followed by chapters on piezoelectric immunosensors and surface plasmon resonance biosensors. A chapter on using nanopores as a label-free biosensing platform and its potential for immunosensing is also included in the second section. The third section starts with a chapter that describes different platforms for carrying out multiplexed immunoassays. This is followed by a chapter on the advantages and limitations of multiplexed bead immunoassays. The Editors express their thanks and appreciation to the authors for their contributions to this book project. Moreover, they are thankful to the Editorial Office at InTech for their support. They are also grateful to the love and support from their families, and acknowledge the assistance from their co-workers. Last but not least, they wish to thank all their former teachers and mentors for sharing their knowledge and experience with them. Dr. Norman H. L. Chiu Department of Chemistry and Biochemistry, The University of North Carolina at Greensboro, USA Dr. Theodore K. Christopoulos Department of Chemistry, University of Patras, Greece [...]... the antigen binding site of heavy chain antibodies is formed only by a single domain that is linked directly via a hinge region to the Fc domain Intact IgG molecules, the bivalent (Fab‘)2, or the monovalent (Fab), all of which contain the antigen binding site(s), can be used in immunoassays Recombinant antibody forms have also been developed to facilitate antibody engineering The single chain fragment... on the binding affinity and specificity of an antibody for its small ligands The affinity maturation processes reported for anti-hapten scFvs are listed in table 4 14 Advances in Immunoassay Technology 6.2.2 Site-directed mutagenesis In site-directed mutagenesis, the investigator changes specific amino acid residues Sitedirected mutagenesis is often used in combination with in silico modeling, crystallographic... Affinity maturation of human botulinum neurotoxin antibodies by light chain shuffling via yeast mating Protein Eng Des Sel, 23(4), 311-319 Makvandi-Nejad, S., Fjallman, T., Arbabi-Ghahroudi, M., Mackenzie, C R., & Hall, J C (2011) Selection and expression of recombinant single domain antibodies from a hyper-immunized library against the hapten azoxystrobin J Immunol Methods in press 20 Advances in Immunoassay. .. antibody’s affinity for its antigen is dependent on the identity and conformation of the amino acid sidechains in the CDRs of both the HC and LC Improvement in the antigen-binding affinity can be attained using a number of strategies The mostly common used are random mutagenesis, site-direct mutagenesis and chain shuffling These processes are often referred to as in vitro affinity maturation, to distinguish... animals or naturally infected animals or humans These libraries are biased for binding to the antigen Thus, affinity maturation takes place in vivo and the chances of isolating the high–affinity antibodies are increased Immune libraries are 6 Advances in Immunoassay Technology constructed using HC and LC variable domain gene pools amplified directly from immune sources; lymphoid sources include peripheral... data, and ligand docking programs, which allow the investigator to hypothesize about the roles that individual binding site amino acid residues have in antigen binding The CDRs of VH and VL are usually targeted for both haptens and protein antigens (Siegel, et al., 2008), and mutations in CDRs as opposed to within the framework residues generally contribute more to increases in affinity (Orcutt et al.,... approaches have been utilized, including identifying the key binding residues, developing more effective procedures for selection of the most specific binders and avoiding interfacial effects that can compromise the yield and stability of rAbs Target Hapten Ab format Antibody library In vitro display Reference Aflatoxin B1 Digoxigenin Doxorubicin Estradiol Indole-3-acetic acid Fluorescein Phenyloxazolone Picloram... obtained by immunization 6 Improving the specificity and affinity of rAbs to low molecular weight ligands Although recombinant antibody technology has been able to open the bottleneck in the isolation of antibodies against virtually any antigen, it remains difficult to obtain high- Recombinant Antibodies and Non-Antibody Scaffolds for Immunoassays 11 affinity antibodies against small molecules using... development of non-biological alternatives to antibodies, including both scaffold proteins and plastibodies, may create distinct opportunities for future improvements in immunoassay technology This could be particularly relevant in applications where compatibility of the binding probe with organic solvents and the ability to withstand 16 Advances in Immunoassay Technology thermal and mechanical stress are required... the binding site This can be a natural binding site or created de novo Randomizations made to the binding region should be able to generate binders with high specificity and affinity (Binz & Pluckthun, 2005b; Gebauer et al., 2009; Gronwall & Stahl, 2009; Kim et al., 2009; Lofblom et al., 2010) All of the scaffolds reported to date, including affibodies, anticalins, and designed ankyrin repeats (DARPin), . of which contain the antigen binding site(s), can be used in immunoassays. Recombinant antibody forms have also been developed to facilitate antibody engineering. The single chain fragment variable. immunoassays have continued to grow exponentially. This book focuses on some of the latest advances in immunoassay technology, which include new materials and methods. The book contains nine. affinity within inexpensive and relatively simple host systems. Effective in vitro libraries have been constructed using either the entire antigen-binding fragment (Fab) or the single chain