Frontiers in Biosensorics I: Fundamental Aspects PDF
Preview Frontiers in Biosensorics I: Fundamental Aspects
EXS 80 Frontiers in Biosensorics I Fundamental Aspects Edited by F. W Scheller F. Schubert J. Fedrowitz Birkhauser Verlag Basel· Boston· Berlin Editors Prof. Dr. EW. Scheller Dr. E Schubert Institut fUr Biochemie Physikalisch-Technische Bundesanstalt und Molekulare Physiologie Abbestrasse 2 -12 c/o Max-Delbriick-Center D-10587 Berlin fUr Molekulare Medizin Robert-Rossle-Strasse 10 D-13122 Berlin Dr. J. Fedrowitz c/o Centrum fUr Hochschulentwicklung POBox 105 D-33311 Giitersloh Library of Congress Cataloging-in-Publication Data A CIP catalogue record for this book is available from the library of Congress, Washington D.C., USA Deutsche Bibliothek Cataloging-in-Publication Data EXS. - Basel; Boston; Berlin: Birkiiuser. Friiher Schriftenreihe Fortlaufende Bei!. zu: Experientia 80. Frontiers in Biosensorics I. Fundamental aspects. - 1997 Frontiers in Biosensorics 1 ed. by. E W. Scheller ... - Basel; Boston; Berlin: Birkhiiuser. (EXS; 80) ISBN -13: 978-3-0348-9883-6 NE: Scheller, Frieder [Hrsg.] I Fundamental aspects. - 1996 Fundamental aspects. - Basel; Boston; Berlin: Birkiiuser. 1997 (Frontiers in Biosensorics I) (EXS; 80) ISBN -13: 978-3-0348-9883-6 The publisher and editor can give no guarantee for the information on drug dosage and administration contained in this publication. The respective user must check its accuracy by consulting other sources of reference in each individual case. The use of registered names, trademarks etc. in this publication, even if not identified as such, does not imply that they are exempt from the relevant protective laws and regulations or free for general use. This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, re-use of illustrations, recitation, broadcasting, reproduction on microfilms or in other ways, and storage in data banks. For any kind of use permission of the copyright owner must be obtained. © 1997 BirkhiiuserVerlag, PO Box 133, CH-4010 Basel, Switzerland Softcover reprint of the hardcover 1st edition 1997 Printed on acid-free paper produced from chlorine-free pulp. TCF 00 ISBN-13: 978-3-0348-9883-6 e-ISBN-J3:978-3-0348 -9043-4 DOl: 10.1007/978-3-0348-9043-4 987654321 Contents F W Scheller, F Schubert and J Fedrowitz Present state and frontiers in biosensorics New recognition elements G. Wulff Imprinting techniques in synthetic polymers - new options for chemosensors ..................... . l3 U E. Spichiger Biomimetic recognition elements for sensor applications 27 W Hummel Screening and characterization of new enzymes for biosensing and analytics . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 M G. Peter and U Wollenberger Phenol-oxidizing enzymes: mechanisms and applications in biosensors ...................... . 63 W F M Stocklein and F W Scheller Enzymes and antibodies in organic media: analytical applications 83 F F Bier and J P Fiirste Nucleic acid based sensors 97 M Sugawara, H. Sato, T. Ozawa and Y. Umezawa Receptor based chemical sensing . . . . . . . . . 121 J Klein, J Altenbuchner and R. Mattes Genetically modified Escherichia coli for colorimetric detection of inorganic and organic Hg compounds . . . . . . . . . . . . . l33 Thin layers/Interfaces L. Brehmer, Y. Kaminorz, R. Dietel, G. Grasnick and G. Herkner Studies on pyroelectric response of polymers modified with azobenzene moieties . . . . . . . . . . . . . . . . . . . . . .. 155 H. Mohwald and R. v. Klitzing Polyelectrolyte layer systems 167 VI Contents M VolkerandH-U Siegmund Forster energy transfer in ultrathin polymer layers as a basis for biosensors . . . . . . . . . . . . . . 175 M Senda Coupling of enzyme reactions to the charge transfer at the interface of two immiscible solvents . . . . . 193 S. Dong and T Chen Electrochemistry of heme proteins on organic molecule modified electrodes . . . . . . . . . . . . . . . . . . . 209 I Katakis and A. Heller Electron transfer via redox hydrogels between electrodes and enzymes . . . . . . . . . . . . . . . . . . . . . . . . 229 T Ikeda Direct redox communication between enzymes and electrodes 243 R. Hintsche, M Paeschke, A. Uhlig and R. Seitz Microbiosensors using electrodes made in Si-technology 267 Subject index 269 Frontiers in Biosensorics I Fundamental Aspects ed. by F. W. Scheller, F. Schubert and J. Fedrowitz © 1997 BirkhauserVeriag Basel/Switzerland Present state and frontiers in biosensorics F. W Scheller I, F. Schubert2 and J. Fedrowitz3 1 Analytcal Biochemistry, Institute of Biochemistry and Molecular Physiology, University of Potsdam, c/o Max-Delbriick-Center ofM olecular Medicine, D-13122 Berlin Germany; 2 c/o CHE Center jor Higher Education Development, D-33311 Giitersloh, Germany; 3 Physikalisch-Technische Bundesanstalt, D-J0587 Berlin, Germany The field of bioanalytics Nature offers an arsenal of interesting principles for the optimization of existing and the realization of novel technical processes. These principles include "classical" reactions of enzymes, antibodies, receptors or nonpro tein macromolecules like nucleic acids, carbohydrates, DNA or RNA. This volume aims at indicating the frontiers in biosensorics by summarizing new recognition elements, new developments in thin layers and interfaces as well as by giving examples for recent applications ofbiosensors. Molecular recognition As far as analysis is concerned, enzymes and antibodies represent powerful tools that allow for sensitive and specific methods of detection and quanti tation to be developed for a wide variety of substances. The first step of the interaction between enzyme and substrate, or anti body and antigen, is the binding of the analyte to the complementary pro tein structure. The basic principle behind the high chemical selectivity of these biomacromolecules is the structural complementarity of the recogni tion elements and the target analyte. While the binding to the antibody normally does not lead to a chemical alteration of the antigen the formation of a complex between substrate and enzyme protein initiates the chemical conversion of the substrate. However, a strict differentiation between both reaction types is not very suitable since they merge into each other. In the absence of cosubstrate an enzyme acts as a binding protein for the substrate; the same is generally true for the binding of enzyme inhibitors and other effectors. Recently catalytic antibodies have been developed which are capable not only of binding the partner but also of catalyzing its chemical converSIOn. Chemoreceptors located in biological membranes function like biomole cular devices driven by the cell metabolism and controlled by the presence 2 F. W. Scheller et al. of biologically active substances. They are extremely attractive tools for the selective recognition of toxins, hormones and drugs. Binding of the agonist either by the opening of ion channels (e.g. with the nicotinic receptor) or via enzyme cascades (e. g. in case of the f3-adrenergic receptor) triggers a signal amplification of 4 to 9 orders of magnitude. The interrogation of the receptor function requires its integration into an artificial membrane; in other cases it can be performed by using (intact) chemosensing structures, e. g. antennae of crustaceans. In addition to the proteic macromolecules nucleic acids and carbohy drates are increasingly used in specialized areas, for example for the sequencing of genomes and for cell surface characterization. The stereo specific interactions of nucleotides - the building blocks of the nucleic acids - are primary for replication of DNA, synthesis of RNA in the tran scription, and ribosomal protein synthesis. Structurally complementary single strands associate to double helical segments where the strength of interaction is determined by the degree of sequence homology. Nowadays, hybridization assays based on this interaction are a fundamental tool of molecular biology and gene identification. Measurement and sequence analysis of DNA and RNA are not only the keys to analyzing the human genome but are unique instruments for genetic profiling susceptibility to diseases, prenatal diagnosis of genetic diseases, and research in molecular biology. -1 : , , -3 , =: , , 0E -5 i , I>'"I) ecc0 -7 .~~. ,,'I' " i0:e~i!l IIc0CE"II\ )) ) wco,oo:>: .E uQ) c 0 (.) -9 Ol ~ -11 -,0 ~I -13 Q. 10 molecular weight Figure 1. Specific features of different bioanalytical procedures. Present state and frontiers in biosensorics 3 The biological recognition elements presented so far cover almost all relevant analytes. As a first step in the design of a bioanalytical process one of them has to be chosen that performs the molecular recognition of the analyte of interest. This selection, however, is dictated by size and concen tration of the analyte (Fig. 1). Signal transduction and bioanalytical configurations In order to obtain a quantifiable output, the interaction between analyte and biomolecule has to be transformed from the chemical into a physico chemical signal. The transfer of information from the biochemical domain to human knowledge requires the command of the techniques for characteriza tion of biochemical systems as well as the transducer principles and techno logies. Furthermore, due to the increased use and availability of computers, it is generally desirable to obtain an electrical output. Thus, the electronic domain is common to all modem bioanalytical instruments. The increasing availability of highly purified enzyme preparations and (monoclonal) antibodies induced the development of a wide range of methods based on these biochemical reagents. For their repeated use, as well as that of cells and other biologically active agents, such as receptors, in analytical devices, numerous techniques for fixing them to carrier ma terials have been developed. Immobilization of the protein, particularly of enzymes, brings about a number of further advantages for their application in analytical chemistry: (i) in many cases the protein is stabilized; (ii) the immobilizate may be easily separated from the sample and (iii) the stable and largely constant biomolecule activity makes the enzyme an integral part of the analytical instrument. Thus, application of immobilized enzymes in analytical chemistry has become common. As early as 1956 the principle of the litmus paper used for pH measure ment was adopted to simplity the enzymatic determination of glucose. By impregnating filter paper with the glucose-converting enzymes the "enzy me test strip" was invented. It can be regarded as the predecessor of optical biosensors and, at the same time, initiated the development of the soc aIled dry chemistry. Nowadays, highly sophisticated enzyme and immuno test strips are commercially available for the determination of about 15 low molecular metabolites and drugs as well as the activities of 10 enzymes. In parallel analytical enzyme- and immuno-reactors have been develop ed where the progress of the reaction is indicated in the reactor effluent colorimetric ally or electrochemically. In packed bed reactors the enzyme catalyzed reaction is carried out in a column of 100 )ll-l 0 ml volume filled with tiny particles bearing the immobilized enzyme. In contrast, in open tubular reactors the enzyme is attached in a monolayer to the inner walls. Such reactors permit a higher measuring frequency. An alternative route was opened in the early 60's by Leland C. Clark, the inventor of the Clark oxygen electrode. He arranged the enzyme solution 4 F. W Scheller et al. directly in front of that electrode and avoided the mixing of the enzyme with the bulk solution by covering the reaction compartment with a semi permeable membrane. Thus, a single enzyme preparation could be used for several samples. This measuring arrangement gave birth to a new sensor concept - the biosensor. The first step in the biosensing process is the specific complex formation of the immobilized recognition element with the analyte. The biological part of a biosensor is often submitted to a conformational change in context with the binding of its partner. In nature this effect may immediately be used for transduction and amplification, e.g., in the ion channels of nerve tissue. The effects of interaction between the analyte molecule and the bio logical system are quantified by the transducer and electronic part of the biosensor. As transducers, chemical sensors, i. e., potentiometric, ampero metric and impedimetric electrodes, optical detectors using indicator dyes, as well as physical sensors, such as piezoelectric crystals, thermistors, and plain optical sensors, have been combined with appropriate biocomponents (Tab. 1). In analogy to affinity chromatography, in so-called binding or affinity sensors, dyes, lectins, antibodies, or hormone receptors are being used in matrix-bound form for molecular recognition of enzymes, glyco proteins, antigens, and hormones. The complex formation changes the magnitude of physico-chemical parameters, such as layer thickness, refrac tive index, light absorption, or electrical charge, which may then be in dicated by means of optical sensors, potentiometric electrodes, or field effect transistors. After the measurement the initial state is regenerated by splitting of the complex. On the other hand, the molecular recognition by enzymes, which can also be applied in the form of organelles, micro organisms and tissue slices, is accompanied by the chemical conversion of the analyte to the respective products. Therefore this type of sensor is termed catalytic or metabolism sensor. It usually returns to the inital state when the Table I. Principles of Biosensors I. Bioaffinity sensors 2. Biocatalytic sensors receptor analyte receptor analyte dye protein substrate enzyme } lectin saccharide organelle cofactor { glycoprotein microbe inhibitor enzyme substrate tissue slice activator inhibitor enzyme activity apoenzyme prosthetic group antibody antigen receptor hormone transport system substrate analogue Transducers optoelectronic detectors, field effect transistors, semiconductor electrodes, potentiometric electrodes, amperometric electrodes, thermistors
The list of books you might like
