Drug Targeting Organ-Specific Strategies

XXIV Contents
11.3.4 Modifications of the Physicochemical Properties of the Protein . . . 282
11.4 The Active Drug . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 283
11.5 The Linkage Between Drug and Carrier . . . . . . . . . . . . . . . . . . . . . 285
11.5.1 Intracellular Degradation . . . . . . . . . . . . . . . . . . . . . . . . . 287
11.5.2 Extracellular Degradation . . . . . . . . . . . . . . . . . . . . . . . . 291
11.6 Recombinant DNA Approaches . . . . . . . . . . . . . . . . . . . . . . . . . 292
11.7 Recombinant DNA Expression Systems . . . . . . . . . . . . . . . . . . . . . 292
11.7.1 Heterologous Gene Expression in Escherichia coli . . . . . . . . . . 292
11.7.2 Fungal Expression Systems . . . . . . . . . . . . . . . . . . . . . . . . 293
11.7.3 Baculovirus Expression Systems . . . . . . . . . . . . . . . . . . . . . 294
11.7.4 Stable Transformations of Insect Cells . . . . . . . . . . . . . . . . . 295
11.7.5 Expression Using Mammalian Cells . . . . . . . . . . . . . . . . . . . 295
11.7.6 Expression Systems: Concluding Remarks . . . . . . . . . . . . . . . 295
11.8 Recombinant DNA Constructs . . . . . . . . . . . . . . . . . . . . . . . . . . 296
11.8.1 Antibody-based Constructs . . . . . . . . . . . . . . . . . . . . . . . 296
11.8.2 Receptor-targeted Constructs . . . . . . . . . . . . . . . . . . . . . . 300
11.8.2.1 Cytotoxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . 300
11.8.2.2 Toxin-targeted Constructs . . . . . . . . . . . . . . . . . . . 300
11.8.2.3 TfR-directed Constructs . . . . . . . . . . . . . . . . . . . . 301
11.9 Recombinant Domains as Building Blocks for Drug Targeting Constructs . . 302
11.9.1 Targeting Domain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
11.9.2 Membrane Translocation Domain . . . . . . . . . . . . . . . . . . . . 303
11.9.3 Assembly Domain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303
11.10 Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 304
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 304
12 Use of Human Tissue Slices in Drug Targeting Research
Peter Olinga, Geny M. M. Groothuis . . . . . . . . . . . . . . . . . . . . . . . 309
12.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 309
12.2 Preparation of Liver Slices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 311
12.3 Incubation and Culture of Liver Slices . . . . . . . . . . . . . . . . . . . . . . 312
12.3.1 Incubation Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . 312
12.3.2 Evaluation of Incubation Systems . . . . . . . . . . . . . . . . . . . . 313
12.3.3 Incubation Systems for Human Liver Slices . . . . . . . . . . . . . . 316
12.3.4 Oxygenation and Culture Media for Liver Slice Incubation . . . . . 316
12.3.5 Pre-incubation of Liver Slices . . . . . . . . . . . . . . . . . . . . . . 317
12.4 Viability and Functionality of Liver Slices . . . . . . . . . . . . . . . . . . . . 317
12.5 In Vitro Transport Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 318
12.5.1 Transport in Hepatocytes . . . . . . . . . . . . . . . . . . . . . . . . . 318
12.5.2 Transport in Liver Slices . . . . . . . . . . . . . . . . . . . . . . . . . 319
12.6 The Use of Liver Slices in Drug Targeting Research . . . . . . . . . . . . . . 321
12.6.1 Distribution and Transport of Drug Targeting Devices . . . . . . . . 321
12.7 Efficacy Testing of the Drug Targeting Device in the Liver . . . . . . . . . . . 323