antibody affinity and affinity distributions

204 Pages·2010·4.98 MB·English

ANTIBODY AFFINITY AND AFFINITY DISTRIBUTIONS
DETERMINATION IN ELISA AND BIOSENSOR
CIP-GEGEVENS KONINKLlJKE BIBLIOTHEEK, DEN HAAG
Pathak, Sulbha Sanjeev
Antibody affinity and affinity distributions.
Determination in ELISA and biosensor/
Sulbha Sanjeev Pathak; [ill.: Tar M. van Osl. - Rotterdam:
Erasmus Universiteit, Afdeling Immunologie. - III.
Thesis Rotterdam. - With ref.
ISBN 90-73436-27-3
NUGI743
Subject headings: antibodies; affinity / enzym immunoassay/
biosensor
No part of this thesis may be reproduced or transmitted in any form by any
means, electronic or mechanical, including photocopying, recording or any
information storage and retrieval system, without permission in writing from the
publisher (S.S. Pathak, Department of Immunology, Erasmus University
Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands).
ANTIBODY AFFINITY AND AFFINITY DISTRIBUTIONS
DETERMINATION IN ELISA AND BIOSENSOR
ANTISTOF AFFINITEIT EN AFFINITEITSDISTRIBUTIES
BEPALING IN ELISA EN BIOSENSOR
PROEFSCHRIFT
ter verkrijging van de graad van doctor
aan de Erasmus Universiteit Rotterdam
op gezag van de rector magnificus
Prof. Dr. P.W.C. Akkermans M. A.
en volgens besluit van het College voor Promoties.
De open bare verdediging zal plaatsvinden op
woensdag 20 december 1995 om 11.45 uur.
door
Sulbha Sanjeev Pathak
geboren te Sang Ii, India
PROMOTIE-COMMISSIE:
Promotor: Prof. dr. R. Benner
Co-promotor: dr. H.F.J. Savelkoul
Overige leden: Prof. dr. H.G. van Eijk
Prof. dr. J.J. Haaijman
Prof. dr. H.J. Neijens
IMMUNOLOGIE The studies described in this
thesis were performed at the
ROTTERDAM
Department of Immunology,
Erasmus University Rotterdam.
Publication of this thesis was partly sponsored by Fisons Applied Sensor
Technology, Cambridge, UK
Illustrations: Tar M. van Os
Printing: Ridderprint, Ridderkerk
To raise new questions, new
possibilities, to regard old problems
from a new angle makes a real
adventure in science.
Where change is the only constant
(Einstein)
To Sanjeev and Gauri
Contents
CHAPTER 1 Introduction 9
CHAPTER 2 Determination of concentration and
affinity in ELISA 45
2.1 Quantitation of immunoglobulin concentration
in ELISA 47
Manual of Immunological Methods. I. Lefkovits (Ed), in press
2.2 Terasaki-ELISA for murine IgE. II. Quantitation
of absolute concentration of antigen-specific
and total IgE 75
J. Immunol. Methods 1989; 116:277-285
2.3 Terasaki-ELISA for murine IgE. III. Determination
of absolute concentration and functional affinity
by sequential equilibrium binding analysis 85
J. Immunol. Methods 1989;183:71-81.
2.4 Mixing of two monoclonal Ig E antibodies
increases the functional affinity of the
mixture for the antigen 97
Submitted
2.5 Determination of relative affinity and affinity
distributions in ELISA 107
Submitted
CHAPTER 3 Kinetic interaction analysis with a biosensor 115
3.1 Determination of antibody affinity and affinity
distributions 117
Manual of Immunological Methods. I. Lefkovits (Ed), in press
3.2 Application of a resonant mirror biosensor
to the determination of functional
affinity of monoclonal and polyclonal
antibodies 141
Submitted
3.3 Interleukin-4 increases the pace of affinity
maturation of IgG 1 antibodies 159
Submitted
CHAPTER 4 Discussion 175
SUMMARY 187
SAMENVA TTING 191
ABBREVIATIONS 195
GLOSSARY 197
DANKWOORD 199
CURRICULUM VITAE 201
PUBLICATIONS 202

INTRODUCTION
Introduction
1.1 Humoral immunity: the role of B cells
1.2 Helper T cells in humoral immunity
1.3 Specificity and diversity
1.3.1 The V(D)J recombination
1.3.2 Somatic hypermutation
1.3.3 Class switch recombination
1.4 Memory
1.4.1 Phenotype of memory Band T cells
1.4.2 Mechanisms of immunological memory
1.5 The germinal center
1.6 The aim of the experimental work
1 .7 References
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