Table Of ContentElectrochemical
Sensors in
Immunological
Analysis
Electrochemical
Sensors in
Immunological
Analysis
Edited by
T.T. Ngo
University of California, Irvine
Irvine, California
Springer Science+Business Media, LLC
Library of Congress Cataloging in Publication Data
Electrochemical sensors in immunological analysis.
Includes bibliographies and index.
1. Immunoassay. 2. Electrochemical analysis. I. Ngo, T. T. (That Tjien), 1 944-
[DNLM: 1. Electrochemistry. 2. Immunoassay — methods. QW 570 E38]
QP519.9J42E44 1987 574/.028 87-1411 6
ISBN 978-1-4899-1976-2
ISBN 978-1-4899-1976-2 ISBN 978-1-4899-1974-8 (eBook)
DOI 10.1007/978-1-4899-1974-8
© Springer Science+Business Media New York 1987
Originally published by Plenum Press, New York in 198 7
Softcover reprint of the hardcover 1s t edition 1987
All rights reserved
No part of this book may be reproduced, stored in a retrieval system, or transmitted
in any form or by any means, electronic, mechanical, photocopying, microfilming ,
recording, or otherwise, without written permissio n from the Publisher
Dedicated to the memory of my parents
11?iJ i
Mr. Ngo Nam Seng (191 3-1 984)
and
t
JIL
Mrs. Thjin Hie Foeng r~ (1 91 3-1 986)
PREFACE
The development of radioimmunoassay (RIA) by R.S. Yalow
and S.A. Berson in 1959 opens up a new avenue in ultra
sensitive analysis of trace substances in complex biological
systems. In recognition of the enormous contributions of RIA
to basic research in biology and to routine clinical tests in
laboratory medicine, R.S. Yalow, the co-developer of RIA, was
awarded, in 1977, the Nobel Prize for Medicine and
Physiology. The basic principle of RIA is elegantly simple.
It is based on a specific, competitive binding reaction
between the analyte and the radio-labeled analog of the
analyte for the specific antibody raised to the analyte. The
combination of high specificity and affinity of an antibody
molecule makes it a very versatile analytical reagent capable
of reacting specifically with analytes at a very low
concentration in a complex solution such as serum. The
sensitivity of RIA is provided by using a radioactive tracer.
RIA is a general technique applicable to the analysis of a
wide range of substances provided that the antibodies to
these substances can be obtained. The two essential reagents
in a RIA are (1) the antibody raised to an analyte and (2) a
radio-labeled analyte or its analog. RIA is typically
performed by simply mixing isotopically labeled analytes and
the specific antibody with the sample, and then separating
the radio-labeled and unlabeled analytes (i.e. the analyte to
be measured) that are unbound to the antibody from the
antibody-bound ones and finally counting the radioactivity
associated with either fraction. The radioactivity is
related to the amount of the analyte by reference to a
standard curve.
Recently, there are intense research efforts to simplify
immunoassays and to expand the applications of this
technology into the physician's office, field-use in
environmental, agricultural and veterinary monitoring and
control where the use of radioactive labels is impractical or
economically infeasible. These concerted research efforts
have led to the development of a number of ingenious non
isotopic labels capable of simplifying the assay by replacing
radioisotopic labels. Some non-isotopic labels have
advantages of allowing the development of a separation-free
(homogeneous) assay format, i.e. a physical separation of the
antibody-bound from the unbound analytes is not required.
This is in contrast to the heterogeneous assay which requirs
a separation step at some point in the assay. Potential
health and environmental hazards associated with the use of
radioisotopes and the limited shelf life of reagents labeled
with isotopes have further stimulated wider applications of
vii
these non-isotopic labels in immunoassay systems. The most
widely used-non-isotopic labels are enzymes, fluorescent
molecules and bio-and chemi-luminogenic reagents. These
labels have been measured mostly by spectrophotometric
methods. Many of these labels can also be conveniently and
sensitively measured electrochemically. The advantages of
using electrochemical sensors over spectrophotometers are:
(1) the capability of making measurements in a highly turbid
sample; (2) excellent detection limits and (3) the low cost
and simplicity of electrochemical devices.
The purpose of this volume is to bring together works of
scientists of diverse backgrounds and to focus attention on
concepts, methodology, limitations and future potentials of
electrochemical sensors in immunological analysis. Topics
includes (1) direct reading immunosensor devices; (2)
separation-free (homogeneous) enzyme mediated immunoassays
with electrochemical detections; (3) liposome mediated
electrochemical immunoassays; (4) applications of ion
selective membrane electrodes in immunoassays; (5) the use
electric pulse to accelerate antigen-antibody interactions in
immunoassays and (6) the combined use of a flow system and
an electrochemical. detector to monitor the immunochemical
reaction.
This volume should complement the recently published
"Enzyme-Mediated Immunoassay", edited by T.T. Ngo and H.M.
Lenhoff, 1985, Plenum Press. Both volumes should be useful to
clinical chemists, clinicians, biochemists, analytical
chemists, toxicologists and biologists.
I am grateful to my wife, Ping Ying, for her
understanding, encouragements and patience while I spent
seemingly endless evenings and week-ends working on this
volume. To my brothers and sisters, in Indonesia and Canada,
who suffered with me the loss of our parents within the last
two years, I wish them well and share with them whatever
recognition this work may bring.
December, 1986. T.T. Ngo
vni
CONTENTS
On the Direct Immunochemical Potentiometric
Signal 1
M. Thompson, J.S. Tauskela and U.J. Krull
Selective Antibody Responsive Membrane Electrodes . 19
M.Y. Keating
Electroimmunoassay of PGE2: An Antibody-Sensitive
Electrode Based Competive Protein-Binding
Assay . 35
G.R. Connell and K.M. Sanders
Immuno-Potentiometric Sensors with Antigen
Coated Membrane • 47
S. Suzuki and M. Aizawa
The Opsonized Electrode. 61
H.A.O. Hill and N.J. Walton
Homogeneous Voltammetric Immunoassay of Haptens • 77
S.A. McClintock and W.C. Purdy
Immunoassay by Differential Pulse 'Polarography
and Anodic Stripping Voltammetry 87
M.J. Doyle, K.R. Wehmeyer, W.R. Heineman
and H.B. Halsall
Separation-Free (Homogeneous) Amperometric
Immunoassay Using an Apoenzyme as the Label . · 103
T.T. Ngo
Digoxin Homogeneous Enzyme Immunoassay
with Amperometric Detection . • 117
D.S. Wright, H.B. Halsall and W.R. Heineman
Antibody to an Enzyme as the Modulator in
Separation-Free Enzyme Immunoassays with
Electrochemical Sensor · 131
T.T. Ngo
Immunochemical Determination of Creatine Kinase
Isoenzyme-MB · 145
S.S. Kuan and G.G. Guilbault
ix
Immunosensor Based on Labeled Liposome
Entrapped-Enzymes . · 167
M. Haga
Measurement of Complement Fixation with
Ion Selective Membrane Electrodes • · 179
P. D'Orazio
Antibody-Antigen Precipitin Reaction Monitoring
with an Ion Selective Electrode • • 195
P.W. Alexander
Enzyme-Linked Immunoassay with a Fluoride
Ion Selective Electrode • • 203
P.W. Alexander
Heterogeneous Potentiometric Enzyme Immunoassay
for Antigens and Haptens with Iodide
Selective Electrode • · 211
J.L. Boitieux, G. Desmet and D. Thomas
Enzyme Immunoassay using the Ammonia
Gas-Sensing Electrode • • 239
C.R. Gebauer
Homogeneous and Heterogeneous Enzyme Immunoassays
Monitored with Carbon Dioxide Sensing
Membrane Electrode · 257
C.A. Broyles and G.A. Rechnitz
Enzyme-Linked Immunosorbent Assays Using
Oxygen-Sensing Electrode • 269
M. Aizawa
Bioaffinity Electrochemical Sensor with Preformed
Metastable Ligand-Receptor Complex · 279
M. Aizawa
Electric Pulse Accelerated Immunoassay . • 293
I. Karube and E. Tamiya
Heterogeneous Enzyme Immunoassay with
Amperometric Detection · 309
K.R. Wehmeyer, M.J. Doyle, H.B. Halsall
and W.R. Heineman
Enzyme Immunoassay in Flow Systems with
Electrochemical and Other Detectors • · 327
B. Mattiasson
Use of Immunochemical Separation and Electrochemical
Detection in Flow Injection Analysis of
Lactic Dehydrogenase • 335
S.S. Kuan and G.G. Guilbault
Rapid Subpicomole Enzyme Immunoassay by Flow
Injection with Amperometric Detection • • 343
W.U. de Alwis and G.S. Wilson
x
Contributors 351
Index 355
xi
