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`PFIZER INC.
`
`PFIZER INC.
`Exhibit 1019
`
`Exhibit 1019
`
`
`
`Monoclonal Antibodies:
`Principles and Practice
`
`Production and Application of
`Monoclonal Antibodies in Cell Biology,
`Biochemistry and Immunology
`
`Second Edition
`
`JAMES W. CODING
`
`Department of Pathology and Immunology,
`Monash University, Alfred Hospital,
`Prahran, Victoria, Australia
`
`ACADEMIC PRESS
`Harcourt Brace Javanovich, Publishers
`London Orlando
`
`_-._“_—_‘_-—__~_‘
`
`PFIZER INC.
`Exhibit 1019-01
`
`
`
`ACADEMIC PRESS INC. (LONDON) LTD.
`24/28 Oval Road
`London NW1
`
`United States Edition published by
`ACADEMIC PRESS
`Orlando, Florida 32887
`
`Copyright © 1986 by
`Academic Press Inc. (London) Ltd.
`
`A ll Rig/{his Reserved
`
`No part of this book may be reproduced in any form by photostal, microfilm, or
`any other means. without written permission from the publishers
`
`British Library Cataloguing in Publication Dala
`
`Goding, James W.
`Monoclonal antibodies: principles and
`practice: production and application of
`monoclonal antibodies in cell biology,
`biochemistry and immunology. —— 2nd ed.
`1. Immunoglobulins
`2. Gammopathics,
`Monoclonal
`I. Title
`QR186.7
`599.02’93
`ISBN 0-12-287021-2
`
`PFIZER INC.
`Exhibit 1019-02
`
`
`
`Contents
`
`Preface to Second Edition
`
`List of Abbreviations
`
`Introduction
`
`2 Theory of Monoclonal Antibodies
`The Clonal Selection Theory
`Structure of Antibodies
`
`Properties of Individual lmmunoglobulin Classes
`Biosynthesis and Assembly of lmmunoglobulins
`Antibody Genes
`Control of the Immune Response
`2.7 Myeloma
`Cell Fusion and Selection of Hybrids
`Hybridomas
`2.10 Differences Between Conventional and Monoclonal
`
`Serology
`
`3 Production of Monoclonal Antibodies
`
`Choice of Normal Lymphocyte Donor
`Immunization Protocol
`
`Choice of Myeloma Cell Line for Fusion
`Equipment Required for Fusion and Growth of Hybridomas
`
`vii
`
`XV
`
`42
`
`59
`59
`61
`64
`66
`
`PFIZER INC.
`Exhibit 1019-03
`
`
`
`Preparation of Spleen Cells
`3.5
`Preparation of Myeloma Cells
`3.6
`Preparation of HAT and HT Medium
`3.7
`Fusion Protocol
`3.8
`Early Growth
`3.9
`3.10 Screening Assays
`3.11 Cloning
`3.12 Large-scale Cultures
`
`Storage of Hybrids in Liquid Nitrogen
`3.13
`3.14 Growth of Hybridomas in Animals
`3.15 Mix-ups of Cells
`3.16 Mycoplasma Contamination
`
`Purification, Fragmentation and Isotopic Labelling
`of Monoclonal Antibodies
`4.1
`Determination of Antibody Class
`4.2 Methods Used in Antibody Purification
`4.3
`Purification of Monoclonal lgC
`4.4
`Purification of lgM
`4.5
`Fragmentation of Monoclonal Antibodies
`
`4.6
`4.7
`
`Radiolabelling of Monoclonal Antibodies
`Conditions for Stability and Storage of MonOclonaI Antibodies
`
`5.3
`5.4
`
`Analysis of Antigens Recognized by Monoclonal
`Antibodies
`5.1
`Determination of the Biochemical Nature of the Antigen
`5.2
`Cellular Distribution of Antigens Detected by Monoclonal
`Antibodies
`Radioiodination of Soluble Protein Antigens
`Radiolabelling of Soluble Proteins with Tritium by Reductive
`Methylation
`Radiolabelling of Cellular Proteins
`Solubilization of Membrane Proteins
`Isolation of Radiolabelled Antigens by Immunoprecipitation
`Electrophoretic Methods for Analysis of Protein Antigens
`Topographical Analysis of Proteins by Monoclonal
`Antibodies
`5.10 Use of Antibodies in the Production and Identification of
`
`5.5
`5.6
`5.7
`5.8
`5.9
`
`Cloned DNA Sequences
`
`Affinity Chromatography Using Monoclonal Antibodies
`6.1
`Preparation of Immobilized Antibodies
`62
`Use of Antibody Affinity Columns
`
`67
`68
`70
`71
`74
`76
`89
`91
`
`91
`92
`93
`93
`
`104
`105
`108
`121
`125
`125
`
`133
`135
`
`142
`142
`
`145
`145
`
`153
`154
`161
`165
`171
`
`199
`
`200
`
`219
`220
`227
`
`PFIZER INC.
`Exhibit 1019-04
`
`
`
`Contents
`
`Immunofluorescence
`Principles of lmmunofluorescence
`The Fluorescence-activated Cell Sorter
`
`Conjugation of Antibodies with Fluorochromes
`The Avidin--Biotin System
`Staining Cells with Fluorescent Antibodies
`Nonspecific Fluorescence
`Lack of Staining
`
`8 Generation of Conventional Antibodies
`
`Strategies for the Preparation of Highly Specific Polyclonal
`Antibodies
`
`Preparative SDS-polyacrylamide Gel Electrophoresis
`Immunization
`
`Purification and Fragmentation of Rabbit, Sheep and Goat
`
`Specific Antibodies from Nonspecrfic Antisera
`
`xi
`
`241
`242
`252
`255
`262
`266
`271
`274
`
`28l
`
`286
`
`288
`290
`
`295
`
`301
`
`PFIZER INC.
`Exhibit 1019-05
`
`
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