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New Comprehensive Biochemistry - Vol 38 : Volume 38 - Makrides

New Comprehensive Biochemistry - Vol 38

Volume 38

By: Makrides

Hardcover

Published: 24th October 2003
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The production of proteins in mammalian cells is an important tool in numerous scientific and commercial areas. For example, proteins for human therapy, vaccination or diagnostic applications are typically produced in mammalian cells. Gene cloning, protein engineering, biochemical and biophysical characterization of proteins also require the use of gene expression in mammalian cells. Other applications in widespread use involve screening of libraries of chemical compounds in drug discovery, and the development of cell-based biosensors.



This book presents a state-of-the-art comprehensive coverage of the technical aspects of gene expression in mammalian cells, written by experienced scientists working at the forefront of the field.

Mentioned in PHARMACEUTICAL RESEARCH, Vol. 21, No. 10, 2004

Prefacep. vii
List of Contributorsp. ix
Other Volumes in the Seriesp. xxxix
Why choose mammalian cells for protein production?p. 1
Abbreviationsp. 6
Referencesp. 6
Vectors for gene expression in mammalian cellsp. 9
Introductionp. 9
Transient gene expressionp. 9
Stable gene expressionp. 14
Genetic elements of mammalian expression vectorsp. 15
Selectable markersp. 21
Signal peptides and fusion moietiesp. 21
mRNA and protein stabilityp. 22
Coordinated expression of multiple genesp. 23
Abbreviationsp. 24
Referencesp. 25
Virus-based vectors for gene expression in mammalian cells: Herpes simplex virusp. 27
Introductionp. 27
Herpes Simplex Virus-relevant basic biologyp. 27
Using HSV-1 to make gene therapy vectorsp. 33
Applications of HSV vectors in the nervous systemp. 42
Applications outside the nervous systemp. 48
Conclusionsp. 50
Abbreviationsp. 51
Referencesp. 51
Virus-based vectors for gene expression in mammalian cells: Epstein-Barr virusp. 55
Introductionp. 55
Replication of the EBV genomep. 55
EBV-based vectors for gene expression in cell culturep. 58
EBV-based vectors for gene therapyp. 60
Cell-specific gene expressionp. 64
A case study for the futurep. 66
Conclusionsp. 67
Abbreviationsp. 68
Referencesp. 68
Virus-based vectors for gene expression in mammalian cells: SV40p. 71
Abstractp. 71
Introductionp. 71
The biology of wild type SV40p. 73
Making recombinant SV40 viruses and using them for gene deliveryp. 76
Conclusions and future perspectivesp. 87
Abbreviationsp. 89
Referencesp. 90
Virus-based vectors for gene expression in mammalian cells: Adeno-associated virusp. 93
Biology of adeno-associated virusp. 93
Production of recombinant AAV vectorsp. 97
In vitro and in vivo gene transferp. 99
Immunological aspects of AAV vectorsp. 102
Development of new AAV vectorsp. 103
Abbreviationsp. 105
Referencesp. 106
Virus-based vectors for gene expression in mammalian cells: Adenovirusp. 109
Introductionp. 109
Adenoviral vectorsp. 111
Gene deliveryp. 115
Gene expressionp. 119
Production and analyses of viral particlesp. 121
Conclusionp. 122
Abbreviationsp. 122
Referencesp. 123
Virus-based vectors for gene expression in mammalian cells: Vaccinia virusp. 125
Introductionp. 125
Vaccinia virus molecular biologyp. 125
Construction of recombinant poxvirus vectorsp. 127
Chimeric VV-bacteriophage expression vectorsp. 130
Improved safety of VV vectorsp. 130
Non-vaccinia poxvirus vectorsp. 132
Laboratory and clinical applicationsp. 132
Abbreviationsp. 134
Referencesp. 134
Virus-based vectors for gene expression in mammalian cells: Baculovirusp. 137
Introductionp. 137
Baculoviruses as insect cell expression vectorsp. 137
Molecular biology of virus vector constructionp. 139
Baculoviruses as mammalian cell expression vectorsp. 139
Characteristics of baculovirus-mediated mammalian gene deliveryp. 142
Applications of the baculovirus mammalian gene delivery vectorp. 145
Points to considerp. 146
Abbreviationsp. 148
Referencesp. 148
Virus-based vectors for gene expression in mammalian cells: Coronavirusp. 151
Introductionp. 151
Coronavirus pathogenicityp. 152
Molecular biology of coronavirusp. 153
Helper-dependent expression systemsp. 154
Single genome coronavirus vectorsp. 156
Optimization of transcription levelsp. 162
Modification of coronavirus tropism and virulencep. 164
Expression systems based on arterivirusesp. 164
Conclusionsp. 165
Abbreviationsp. 165
Referencesp. 166
Virus-based vectors for gene expression in mammalian cells: Poliovirusp. 169
Introductionp. 169
A poliovirus vector-based HIV vaccinep. 169
Poliovirus vector development and its immunogenic potentialp. 176
Abbreviationsp. 185
Referencesp. 185
Virus-based vectors for gene expression in mammalian cells: Sindbis virusp. 189
Introductionp. 189
Viral genetic elementsp. 190
Packaging of repliconsp. 192
Effects of alphaviruses and replicons on infected cellsp. 193
Expression in neuronsp. 194
Insect and crustaceap. 195
Vaccines for infectious diseases and cancerp. 198
Perspectivesp. 202
Abbreviationsp. 203
Referencesp. 203
Virus-based vectors for gene expression in mammalian cells: Semliki Forest virusp. 207
Introductionp. 207
Expression of topologically different proteinsp. 209
Host cell rangep. 212
Scale-up of protein productionp. 215
Expression in primary cell culturesp. 216
Expression in hippocampal slice culturesp. 217
SFV vectors in vivop. 219
Safety of SFV vectorsp. 219
SFV vectors in gene therapyp. 220
SFV vectors as tools for virus assemblyp. 221
Modifications of SFV vectorsp. 222
Novel technologiesp. 224
Conclusionsp. 225
Abbreviationsp. 226
Referencesp. 227
Virus-based vectors for gene expression in mammalian cells: Retrovirusp. 231
Introductionp. 231
Biology of retrovirusesp. 231
Development of recombinant retrovirus vectorsp. 235
Expression of the transgenep. 242
Targetingp. 245
Conclusion and perspectivesp. 247
Abbreviationsp. 247
Referencesp. 248
Virus-based vectors for gene expression in mammalian cells: Lentivirusesp. 251
Introductionp. 251
Genetic structure and biology of lentivirusesp. 251
HIV-1-derived vector packaging systemp. 255
Lentiviral vector productionp. 258
Pseudotyped vectorsp. 259
Expression from lentiviral vectorsp. 260
Gene transfer applicationsp. 262
Conclusionp. 262
Abbreviationsp. 262
Referencesp. 263
Methods for DNA introduction into mammalian cellsp. 265
Introductionp. 265
Barriers to successful transfectionp. 266
Comparison of available methodsp. 269
Applicationsp. 274
Abbreviationsp. 275
Referencesp. 276
Lipid reagents for DNA transfer into mammalian cellsp. 279
Introductionp. 279
General structure of cationic lipidsp. 279
Formulation and physicochemical propertiesp. 280
Mechanisms of in vitro transfectionp. 282
In vivo administrationp. 284
Targetingp. 285
New strategies to improve in vivo transfectionp. 286
Conclusionp. 286
Abbreviationsp. 287
Referencesp. 287
Reporter genes for monitoring gene expression in mammalian cellsp. 291
Introductionp. 291
Reporter gene vectors and fusionsp. 292
Green fluorescent proteinp. 293
Luciferasesp. 299
Alkaline phosphatasep. 303
Chloramphenicol acetyltransferasep. 304
[beta]-Galactosidasep. 305
Concluding remarksp. 306
Abbreviationsp. 307
Referencesp. 307
Gene transfer and gene amplification in mammalian cellsp. 309
Introduction on the origin of Chinese hamster ovary cells for recombinant protein productionp. 309
The DHFR/methotrexate/CHO expression system: a multi-layer selection system for high-level expression of recombinant genesp. 311
Gene amplification is a phenomenon occurring frequently in immortalized mammalian cellsp. 316
Concluding remarksp. 330
Abbreviationsp. 332
Referencesp. 333
Co-transfer of multiple plasmids/viruses as an attractive method to introduce several genes in mammalian cellsp. 337
Introductionp. 337
Multiplicity of infection (MOI) and multiplicity of transfection (MOT)p. 337
Random distribution of vector unitsp. 339
Several copies of one genep. 340
Co-transfer of multiple genesp. 343
Conclusionp. 346
Abbreviationsp. 347
Referencesp. 347
Optimization of plasmid backbone for gene expression in mammalian cells
Introductionp. 349
Bacterial DNA sequencesp. 349
Safetyp. 351
Plasmid sizep. 351
Optimization of plasmids for nuclear localizationp. 352
Examples of vectors: pCOR and minicirclesp. 354
Conclusionsp. 355
Abbreviationsp. 356
Referencesp. 356
Use of scaffold/matrix-attachment regions for protein productionp. 359
Introductionp. 359
Chromatin structure and control of gene expressionp. 360
Use of MAR elements for protein productionp. 370
Conclusionp. 376
Abbreviationsp. 376
Referencesp. 377
Chromatin insulators and position effectsp. 381
Introductionp. 381
Chromosomal position effectsp. 382
Chromatin insulatorsp. 387
Specific uses of chromatin insulatorsp. 390
Abbreviationsp. 393
Referencesp. 394
Locus Control Regionsp. 397
Introductionp. 397
The [beta]-globin locus LCRp. 398
Properties of LCRsp. 401
LCR knockout micep. 403
Mechanisms of globin gene activation by the LCRp. 404
LCRs of other genesp. 405
Abbreviationsp. 406
Referencesp. 406
Protein synthesis, folding, modification, and secretion in mammalian cellsp. 411
Introductionp. 411
Modification of proteins in the early secretory pathwayp. 412
Golgi modificationsp. 418
Quality controlp. 421
Transport of proteins through the secretory pathwayp. 424
Conclusionp. 428
Abbreviationsp. 428
Referencesp. 429
Pathways and functions of mammalian protein glycosylationp. 433
Introductionp. 433
Glycosylation of asparagine residues: N-glycosylationp. 434
Glycosylation of serine/threonine residues: O-glycosylationp. 440
Glycosyl phosphatidyl inositol (GPI)-anchored proteinsp. 444
What specifies glycan structure?p. 447
Functions of protein-linked glycansp. 449
Abbreviationsp. 452
Referencesp. 453
Metabolic engineering of mammalian cells for higher protein yieldp. 457
Therapeutic protein production from mammalian cellsp. 457
Initial metabolic engineering approachesp. 458
Transcriptional hotspot targeting, chromosomal locus amplification and episomal expression strategiesp. 458
Biphasic production processes--controlled proliferation in biotechnologyp. 459
Counteracting suicidal tendencies: preventing cell death in mammalian cell culturesp. 463
Third generation metabolic engineering--multiregulated multigene metabolic engineeringp. 465
Outlookp. 466
Abbreviationsp. 467
Referencesp. 467
Translational regulation in mammalian cellsp. 471
Introductionp. 471
Overview of the initiation phase of translation in eukaryotesp. 471
Regulation of initiation via mRNA structurep. 476
Regulation via initiation factorsp. 481
Regulation of initiation via mRNA-specific binding proteinsp. 484
Regulation of translation subsequent to the initiation stepp. 486
Complicated issues and closing notesp. 487
Abbreviationsp. 491
Referencesp. 491
Pathways of mammalian messenger RNA degradationp. 495
Summaryp. 495
mRNA decay comprises a major control in gene expressionp. 495
General pathways of mRNA decayp. 496
Regulation of mRNA decay by rate-limiting endonuclease cleavagep. 499
Decay pathways involved in mRNA surveillancep. 500
Examples of decay pathways controlled by defined cis-trans interactionsp. 504
The role of translation in mRNA decay pathwaysp. 509
Degradation of mRNA by external factorsp. 509
Conclusionsp. 510
Abbreviationsp. 510
Referencesp. 511
Pathways of mammalian protein degradationp. 513
Introductionp. 513
Pathways of selective proteolysisp. 513
Pathways of nonselective proteolysisp. 524
Summaryp. 531
Abbreviationsp. 531
Referencesp. 532
Stabilization of proteasomal substrates by viral repeats
Introductionp. 535
The ubiquitin--proteasome systemp. 535
Viral repeats that block proteasomal degradationp. 537
Possible applications in gene transfer settingsp. 544
Concluding remarksp. 547
Abbreviationsp. 547
Referencesp. 547
Architecture and utilization of highly expressed genomic sitesp. 551
Introductionp. 551
Chromatin domains and transgene integration targetsp. 551
Comparison between gene transfer by transfection and retroviral infectionp. 558
Re-use of established highly expressed genomic sitesp. 563
Lessonsp. 569
Abbreviationsp. 570
Referencesp. 571
Intracellular targeting of antibodies in mammalian cells
Introductionp. 573
Concept of intrabodies and their mechanism of actionp. 573
Critical parameters for a successful sFv intrabodyp. 575
The source and selection of sFv antibody for intrabody constructionp. 576
Recent applicationsp. 579
Future perspectivesp. 584
Abbreviationsp. 585
Referencesp. 585
Inducible gene expression in mammalian cellsp. 589
Introductionp. 589
Artificial transcription control systemsp. 590
Generic strategies to improve key characteristics of basic gene regulation systemsp. 597
Complex regulation systemsp. 598
Conclusionsp. 602
Abbreviationsp. 602
Referencesp. 603
Protein production by large-scale mammalian cell culturep. 605
Introductionp. 605
Cell Linesp. 606
Cell culture media and raw materialsp. 607
Cell expansion processp. 609
Cell culture process development and optimizationp. 611
Large scale cell culture process choicesp. 612
Process scale-up challengesp. 615
Large scale bioreactor designp. 617
Process validationp. 619
Conclusionp. 620
Abbreviationsp. 620
Referencesp. 621
Protein production in transgenic animalsp. 625
Introductionp. 625
Production of recombinant proteins in the milk of transgenic animalsp. 626
Other transgenic bioreactorsp. 634
Conclusionsp. 637
Referencesp. 637
Strategies for the purification of recombinant proteinsp. 641
Introductionp. 641
Developing a rational approach for protein purificationp. 641
Assay developmentp. 642
Product recovery and initial purificationp. 643
Purification of proteins from culture mediap. 644
Purification of proteins from cell lysatesp. 644
Volume reduction and partial purificationp. 645
Chromatographic purification of recombinant proteinsp. 649
Product stability and long-term storagep. 657
Abbreviationsp. 658
Referencesp. 658
Indexp. 661
Table of Contents provided by Rittenhouse. All Rights Reserved.

ISBN: 9780444513717
ISBN-10: 044451371X
Series: New Comprehensive Biochemistry
Audience: Professional
Format: Hardcover
Language: English
Number Of Pages: 722
Published: 24th October 2003
Publisher: Elsevier Science & Technology
Country of Publication: GB
Dimensions (cm): 21.08 x 16.49  x 5.61
Weight (kg): 1.58