Characterization of Nanoparticles Intended for Drug Delivery (Methods in Molecular Biology, 2789)

This third edition volume expands on the previous editions with new and updated discussions on the latest developments in endotoxin contamination, complex physicochemical properties, in vitro immunotoxicity traits, and in vitro drug release properties. Eight chapters in this book are dedicated to physicochemical characterization techniques and cover newer methods such as asymmetric-flow field-flow fractionation, single particle inductively coupled plasma mass spectrometry, and resistive pulse sensing. The next eighteen chapters explore the immunotoxicity of nanomaterials, including microbial contaminants such as endotoxin and beta-glucans, anti-PEG antibodies, autoimmunity, and immunosuppressive properties. The last two chapters talk about new pharmacology protocols, including a new technique to assess drug release and a tissue distribution assay using PEG immunohistochemistry. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step instructions to reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Characterization of Nanoparticles Intended for Drug Delivery, Third Edition is a valuable resource that will help both expert and novice researchers further enhance their understanding of this important and developing field. Preface Contents Contributors Part I: Introduction Chapter 1: Advancements in Nanoparticle Characterization 1 Introduction 2 Physicochemical Characterization 3 Immunology 4 Pharmacology and Toxicology 5 The Future of Characterization References Part II: Physicochemical Characterization Chapter 2: Nanoparticle Size Distribution and Stability Assessment Using Asymmetric-Flow Field-Flow Fractionation 1 Introduction 2 Materials 3 Methods 3.1 AF4 Method Optimization 3.2 System Setup 3.3 Conditioning the New Membrane 3.4 Sample Preparation 3.5 Data Analysis 4 Notes References Chapter 3: Assessment of Protein Binding Using Asymmetric-Flow Field-Flow Fractionation Combined with Multi-angle Light Scatte... 1 Introduction 2 Materials 3 Methods 3.1 AF4 Method Optimization, System Setup, and Membrane Conditioning 3.2 Sample Preparation 3.3 Data Analysis 4 Notes References Chapter 4: Quantitation of Active Pharmaceutical Ingredients in Polymeric Drug Products Using Elemental Analysis 1 Introduction 2 Materials 3 Methods 3.1 Instrument Preparation 3.2 Preparation of Standards and Sample 3.3 Stabilization and Calibration 3.4 Sample Analysis 3.5 Shut Down 3.6 Data Analysis 4 Notes References Chapter 5: Particle Size and Concentration Measurement Using the Spectradyne nCS1 Instrument 1 Introduction 2 Materials 3 Methods 3.1 Sample Measurement 3.2 Shutdown and Cleaning Procedure 3.3 Data Analysis 4 Notes References Chapter 6: Measuring Size and Number Concentration of Metallic Nanoparticles Using spICP-MS 1 Introduction 2 Materials 3 Methods 3.1 Measuring the Flow Rate of the Peristaltic Pump 3.2 Preparation of Dissolved Elemental Standards (Au Standards) 3.3 Prepare Nanoparticle Standard Solutions and Unknown Samples 3.4 Sample Measurement 3.5 Data Analysis 3.5.1 Processes for Determining the Sizes for the Unknown Particles, Method 1: Nanoparticle Calibration Curve (See Note 22) 3.5.2 Processes for Determining the Sizes for the Unknown Particles, Method 2: Using the Dissolved Standards Curve and the Den... 3.5.3 Determine the Particle Concentration of the Unknown Nanoparticle Solution 4 Notes References Chapter 7: Ion Quantitation in Liposomal Products Using RP-HPLC with Charged Aerosol Detection 1 Introduction 2 Materials 3 Methods 3.1 Total Ammonium Ion Concentration 3.2 External Ammonium Ion Concentration 3.3 Total Sulfate Ion Concentration 3.4 External Sulfate Ion Concentration 3.5 Data Analysis 4 Notes References Chapter 8: A Static Headspace Gas Chromatography Method for Quantitation of Residual Solvents in Nanoformulations 1 Introduction 2 Materials 3 Method 3.1 Method Validation 3.2 Calculations 3.3 Procedure for Shutting Down the GC 4 Notes References Part III: Immunotoxicity Chapter 9: Current Considerations and Practical Solutions for Overcoming Nanoparticle Interference with LAL Assays and Minimiz... 1 Introduction 2 Choosing an LAL Format 3 Overcoming False-Positive Results Due to Contamination with Beta-Glucans 4 Overcoming False-Negative Results Due to the Presence of Cationic Charge 5 Overcoming False-Negative Results Due to Endotoxin Entrapment 6 Overcoming False-Negative Results Due to Endotoxin Binding to Proteins and Containers 7 The Use of Commercial Dispersion Reagents 8 Some Experimental Approaches to Remove Endotoxin from Reagents, Equipment, Contaminated Nanoparticle Formulations, and Their... 9 Precautions to Avoid Endotoxin Contamination in Nanoformulations 10 Notes References Chapter 10: Detection of Beta-Glucan Contamination in Nanoparticle Formulations 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Reagents 3.2 Preparation of Standards, Controls, and Test Samples 3.3 Experimental Procedure 3.4 Data Report and Interpretation 3.5 Acceptance Criteria 4 Notes References Chapter 11: Detection of Intracellular Complement Activation by Nanoparticles in Human T Lymphocytes 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Reagents and Controls 3.2 Preparation of Test Nanomaterials 3.3 PBMC Isolation 3.4 Procedure for Cell Treatment with Nanoparticles and Controls 3.5 Preparation of Cells for Flow Cytometry 3.6 Preparation of Instrument for Flow Cytometry 3.7 Data Analysis and Report 3.8 Acceptance Criteria 4 Notes References Chapter 12: Analysis of Nanoparticles ́ Potential to Induce Autoimmunity 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Study Samples and Controls 3.2 Experimental Procedure 3.3 Data Analysis and Interpretation 4 Notes References Chapter 13: Analysis of Nanoparticles ́ Effects on Drug-Induced Psoriasis 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Study Samples and Controls 3.2 Experimental Procedure 3.2.1 Prophylactic Mode 3.2.2 Therapeutic Mode 3.3 Data Analysis and Interpretation 4 Notes References Chapter 14: Detection of Nanoparticle-Mediated Total Oxidative Stress in T Cells Using CM-H2DCFDA Dye 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Study Samples and Controls 3.2 Preparation of T Cells 3.3 Experimental Procedure 3.4 Data Analysis and Acceptance Criteria 4 Notes References Chapter 15: Detection of Induction of Mitochondrial Oxidative Stress by Nanoparticles in T Cells Using MitoSOX Red Dye 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Study Samples and Controls 3.2 Preparation of T Cells 3.3 Experimental Procedure 3.4 Data Interpretation 4 Notes References Chapter 16: Detection of Nanoparticle-Mediated Change in Mitochondrial Membrane Potential in T Cells Using JC-1 Dye 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Study Samples and Controls 3.2 Collection and Handling of Whole Blood for Culture 3.3 Isolation of T Cells 3.4 Experimental Procedure 3.5 Data Interpretation 4 Notes References Chapter 17: Detection of Antigen Presentation by Murine Bone Marrow-Derived Dendritic Cells After Treatment with Nanoparticles 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Study Samples and Controls 3.2 Isolation and Counting Bone Marrow Cells 3.3 Generation of Immature Dendritic Cells 3.4 Experimental Procedure 3.5 Data Analysis and Report 4 Notes References Chapter 18: Antigen-Specific Stimulation of CD8+ T Cells by Murine Bone Marrow-Derived Dendritic Cells After Treatment with Na... 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Study Samples and Controls 3.2 Isolation and Counting Bone Marrow Cells 3.3 Generation of Immature Dendritic Cells 3.4 Treatment with Nanoparticles and Controls 3.5 Extraction and Homogenization of OT-I Spleen 3.6 CD8+ T Cell Labeling and Purification 3.7 Setting Up Cell Stimulation Experiment 3.8 Assessing T Cell Purity After Isolation 3.9 Ending Experiment 3.10 Data Analysis and Report 4 Notes References Chapter 19: Detection of Pre-Existing Antibodies to Polyethylene Glycol and PEGylated Liposomes in Human Serum 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Reagents and Controls 3.2 Experimental Procedure 3.3 Data Analysis and Interpretation 4 Notes References Chapter 20: In Vitro and In Vivo Methods for Analysis of Nanoparticles' Potential to Induce Delayed-Type Hypersensitivity Reac... 1 Introduction 2 Materials (See Note 1) 2.1 hCLAT and MUSST 2.2 hCLAT Only 2.3 MUSST Only 2.4 LLNP and LLNA 3 Methods 3.1 Preparation of Controls and Cell Lines 3.2 Preparation of Study Samples Box 1 Box 1 Example Calculation of Nanoparticle Concentration for In Vitro Test 3.3 hCLAT Experimental Procedure 3.4 MUSST Experimental Procedure 3.5 LLNA and LLNP Experimental Procedure 3.6 Calculations and Data interpretation: hCLAT and MUSST 3.7 Calculations and Data Interpretation: LLNA and LLNP 4 Notes References Chapter 21: Analysis of Nanoparticle Adjuvant Properties 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Control, Test Antigen, and Test Nanoparticle 3.2 In Vivo Assay 3.3 Calculations and Result Interpretation 4 Notes References Chapter 22: Methods for Analysis of Nanoparticle Immunosuppressive Properties 1 Introduction 2 Materials 2.1 HuLA 2.2 TDAR 3 Methods 3.1 Preparation of Controls and Study Sample for HuLA Box 1: Example Calculation of Nanoparticle Concentration for In Vitro Test 3.2 Preparation of Study Sample for TDAR 3.3 Human Lymphocyte Activation (HuLA) Assay 3.4 TDAR Assay 3.5 Calculations and Data Interpretation for HuLA 3.6 Calculations and Data Interpretation for TDAR 4 Notes References Chapter 23: Evaluation of the Acute Anaphylactoid Reactogenicity of Nanoparticle-Containing Medicines and Vaccines Using the P... 1 Introduction 2 Materials 3 Methods 3.1 Animal Preparation and Surgical Procedure 3.2 Drug Dosing and Animal Monitoring 3.3 Analysis of Collected Blood Samples 3.4 Data Analysis and Reporting 4 Notes References Chapter 24: Immunophenotyping, Part I: Instrument Calibration and Reagent Qualification for Immunophenotyping Analysis of Huma... 1 Introduction 2 Materials 2.1 Reagents for PBMC Cultures 2.2 Reagents for Flow Cytometry 2.3 General Supplies and Equipment 3 Methods 3.1 Preparation of Reagents and Controls 3.2 PBMC Isolation 3.3 Optional Antibody Titration for Flow Cytometry (see Notes 18-20) 3.3.1 Bead and Live/Dead PBMC Plate Preparation (Without Antibodies or Dyes) 3.3.2 Antibody/Dye Titration Plate Preparation 3.3.3 Combination of Antibody/Dye Titrations to Bead and Live/Dead PBMC Plate 3.3.4 Plate Reading with NovoCyte 3005 3.3.5 Data Analysis in NovoExpress 3.4 Single Stain Controls with Compensation Beads (see Notes 18 and 33) 3.4.1 Plate Preparation with Beads and Cells 3.4.2 Antibody Dilution Preparation 3.4.3 Plate Reading 3.5 Flow Cytometry Control Experiments with PBMC and FMO Controls: Day 1, PBMC (see Note 40) 3.6 Flow Cytometry Control Experiments with PBMC and FMO Controls: Day 2, FMO Controls (see Note 40) 3.6.1 Prepare Heat Shocked PBMC 3.6.2 Harvest and Prepare PBMC 3.6.3 Stain Cells 3.6.4 Data Acquisition with NovoCyte 3005 for FMO Controls 3.6.5 Data Analysis of FMO Controls with NovoExpress (see Note 47) 3.6.6 Analysis Method/Compensation Adjustment of FMO Controls 4 Notes References Chapter 25: Immunophenotyping, Part II: Analysis of Nanoparticle Effects on the Composition and Activation Status of Human Per... 1 Introduction 2 Materials 2.1 Reagents for PBMC Cultures 2.2 Reagents for Flow Cytometry 2.3 General Supplies and Equipment 3 Methods 3.1 Preparation of Reagents and Controls 3.2 PBMC Isolation and Treatment with Nanoparticles and Control (Day 1) 3.3 Master Mix Preparation, PBMC Harvesting, and PBMC Staining (Day 2) 3.4 Sample Preparation Before Analysis on a Flow Cytometer (Day 3) 3.5 Data Acquisition on Novocyte 3005 (Day 3) 3.6 Applying Compensation Matrix to Immunophenotyping Panels for Data Analysis (See Note 31) 3.7 Data Analysis of Immunophenotyping Panel 1 3.8 Data Analysis of Immunophenotyping Panel 2 4 Notes References Chapter 26: Understanding the Role of Scavenger Receptor A1 in Nanoparticle Uptake by Murine Macrophages 1 Introduction 2 Materials 3 Methods 3.1 Preparation of Reagents and Controls 3.2 Preparation of Study Samples 3.3 Preparation of Cells 3.4 Experimental Procedure 3.5 Calculations and Data Interpretation 4 Notes References Part IV: Pharmacology Chapter 27: Assessment of Temperature-Dependent Drug Release of Solubilizing Nanoformulations Using the SITUA 1 Introduction 2 Materials 3 Methods 3.1 Temperature-Dependent Drug Release in 4.5% HSA 3.2 LC-MS Set Up 3.3 Calibration and Quality Control Standards Preparation 3.4 Sample Preparation with Controls 3.5 Calculations 4 Notes References Chapter 28: Evaluation of Nanomedicine Tissue Distribution and Stability by Alexa Fluor 488 and PEG Immunohistochemistry 1 Introduction 2 Materials 3 Methods 3.1 Tissue Sample Preparation 3.2 Alexa Fluor 488 Immunohistochemistry (Bench Staining Procedure) 3.3 PEG (Polyethylene Glycol) Immunohistochemistry (Autostainer Procedure) 4 Notes References Index