Monoclonal Antibodies: A Practical Approach

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Philip Shepherd, Christopher J. Dean
Oxford University Press, 2000 - Medical - 479 pages
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Monoclonal Antibodies: A Practical Approach covers the preparation, testing, derivation, and applications of monoclonal antibodies. New immunological techniques incorporating tried and tested methodologies are described, making the book of interest to established and inexperienced immunologists. Both the standard somatic hybridization technique and recombinant techniques, including the use of phage libraries, for the preparation of rodent and human monoclonal antibodies are described. Protocols for both the small and large scale production are detailed, as well as purification and labelling (with both radioisotopes and non-radioisotopes) methods. The applications of monoclonal antibodies in immunoblotting, enzyme linked immunoassays, immunofluorescence, and FACS analysis are all covered in detail. Finally protocols are given for the use of monoclonal antibodies in rheumatoid arthritis, tissue typing, detecting DNA modified during chemotherapy, and in the clinical analysis of transplantation samples for malignancy. This book will therefore be an invaluable laboratory companion to anyone using monoclonal antibodies in their research.
  

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Contents

Preparation of recombinant antibodies from immune rodent
25
Appendix
58
j Selection of antibodies from phage libraries of immunoglobulin
67
Proximity selections
78
ARM complexes for in vitro display and evolution of antibody
91
Ribosome display methodology
93
Troubleshooting
107
EBV transformation of B cells
114
Radiolabelling of monoclonal antibodies
207
Nonradioactive antibody probes
237
Immunogold probes for light and electron microscopy
247
Characterization of cellular antigens using monoclonal antibodies
265
Immunoassays
297
Immunoaffinity chromatography of macromolecules
319
Immunochemical detection of BrdUrd labelled nuclei for monitoring cell
341
Immunofluorescence microscopy
355

Cryopreservation of cells
122
Laboratory based methods for small scale production of monoclonal
125
Stirred cultures
131
Perfusion high cell densityl systems
137
Harvesting and concentration
143
Isolation and purification of monoclonal antibodies from tissue
149
Precipitation methods
156
Antibody production in plants
181
FACS analysis of clinical haematological samples in transplantation
371
Immunocytochemical staining of cells and tissues for diagnostic
391
Detection of chemically modified DNA in lymphocytes of patients undergoing
411
Monoclonal antibody therapy in organ transplantation
431
Monoclonal antibody therapy in rheumatoid arthritis
449
Al List of suppliers
463
index
469
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About the author (2000)

Phil S. Shepherd is at The Guy's, King's College, and St Thomas's Hospitals, London.

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