Series: Methods in Molecular Biology, Vol. 697

McNeil, Scott E. (Ed.)

2011, XII, 269p.

Hardcover

• Provides the reader with protocols that have been tested against clinically relevant nanoparticles
• Describes the nuances of nanoparticle types and necessary controls
• Gives a broad overview of applications of nanotechnology to medicine
• Outlines the beneficial aspects of nanotechnology-based drug delivery

Content Level » Professional/practitioner

Keywords » Assays for sterility - Assays for toxicity - Atomic force microscopy - Nanomedicine - Physicochemical properties - Thin layer chromatography

Contents
Preface . v
Contributors ix

Part I Introduction and Discussion

1 Unique Benefits of Nanotechnology to Drug Delivery and Diagnostics . . . . . . . .     3
Scott E. McNeil

2 Challenges for Nanoparticle Characterization . . . . . . . . . . . . . . . . . . . . . . . . . . . .     9
Scott E. McNeil

3 Considerations When Submitting Nanotherapeutics to FDA/CDER
for Regulatory Review . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     17
Katherine Tyner and Nakissa Sadrieh

Part II A ssays to Determine Physical and Chemical Properties

4 Measuring the Hydrodynamic Size of Nanoparticles in Aqueous
Media Using Batch-Mode Dynamic Light Scattering . . . . . . . . . . . . . . . . . . . . . .     35
Vincent A. Hackley and Jeffrey D. Clogston

5 Characterization of Nanoparticles by Matrix Assisted Laser Desorption
Ionization Time-of-Flight Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . .     53
Uma Ramalinga, Jeffrey D. Clogston, Anil K. Patri, and John T. Simpson

6 Zeta Potential Measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     63
Jeffrey D. Clogston and Anil K. Patri

7 Size Measurement of Nanoparticles Using Atomic Force Microscopy . . . . . . . . . .     71
Jaroslaw Grobelny, Frank W. DelRio, Namboodiri Pradeep,
Doo-In Kim, Vincent A. Hackley, and Robert F. Cook

8 Biological Tissue and Cell Culture Specimen Preparation
for TEM Nanoparticle Characterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     83
Kunio Nagashima, Jiwen Zheng, David Parmiter, and Anil K. Patri

9 SEM X-Ray Microanalysis of Nanoparticles Present in Tissue
or Cultured Cell Thin Sections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     93
Jiwen Zheng, Kunio Nagashima, David Parmiter,
Jason de la Cruz, and Anil K. Patri

10 Detecting and Measuring Free Gadolinium in Nanoparticles
for MRI Imaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     101
Jeffrey D. Clogston and Anil K. Patri

11 Lipid Component Quantitation by Thin Layer Chromatography . . . . . . . . . . . . .     109
Jeffrey D. Clogston and Anil K. Patri

Part III A ssays for Sterility

12 Detection and Quantitative Evaluation of Endotoxin Contamination
in Nanoparticle Formulations by LAL-Based Assays . . . . . . . . . . . . . . . . . . . . . . .     121
Barry W. Neun and Marina A. Dobrovolskaia

13 Analysis of Microbial Contamination in Nanoparticle Formulations . . . . . . . . . . .     131
Timothy M. Potter and Marina A. Dobrovolskaia

Part IV A ssays for In Vitro Detection and Quantitation

14 Gold Nanoparticle Quantitation via Fluorescence in Solution and Cell Culture . .     137
Parag Aggarwal and Marina A. Dobrovolskaia

15 Quantitation of Nanoparticles in Serum Matrix by Capillary Electrophoresis . . . .     145
King C. Chan, Timothy D. Veenstra, and Haleem J. Issaq

Part V In Vitro Assays for Toxicity

16 Evaluation of Cytotoxicity of Nanoparticulate Materials in Porcine
Kidney Cells and Human Hepatocarcinoma Cells . . . . . . . . . . . . . . . . . . . . . . . . . 157
Timothy M. Potter and Stephan T. Stern

17 Monitoring Nanoparticle-Treated Hepatocarcinoma Cells for Apoptosis . . . . . . .     167
Timothy M. Potter and Stephan T. Stern

18 Detecting Reactive Oxygen Species in Primary Hepatocytes
Treated with Nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     173
Banu Zolnik, Timothy M. Potter, and Stephan T. Stern

19 Assay to Detect Lipid Peroxidation upon Exposure to Nanoparticles . . . . . . . . . .     181
Timothy M. Potter, Barry W. Neun, and Stephan T. Stern

20 Monitoring Glutathione Homeostasis in Nanoparticle-Treated Hepatocytes . . . . .     191
Timothy M. Potter, Barry W. Neun, and Stephan T. Stern

21 Autophagy Monitoring Assay: Qualitative Analysis
of MAP LC3-I to II Conversion by Immunoblot . . . . . . . . . . . . . . . . . . . . . . . . .     199
Christopher B. McLeland, Jamie Rodriguez, and Stephan T. Stern

22 Monitoring Lysosomal Activity in Nanoparticle-Treated Cells . . . . . . . . . . . . . . .     207
Barry W. Neun and Stephan T. Stern

Part VI In Vitro Immunological Assays

23 Method for Analysis of Nanoparticle Hemolytic Properties In Vitro . . . . . . . . . . .     215
Barry W. Neun and Marina A. Dobrovolskaia

24 Method for In Vitro Analysis of Nanoparticle Thrombogenic Properties . . . . . . .     225
Barry W. Neun and Marina A. Dobrovolskaia

25 Qualitative Analysis of Total Complement Activation by Nanoparticles . . . . . . . . .     237
Barry W. Neun and Marina A. Dobrovolskaia

26 Method for Analysis of Nanoparticle Effects on Cellular Chemotaxis . . . . . . . . . .     247
Sarah L. Skoczen, Timothy M. Potter, and Marina A. Dobrovolskaia

27 In Vitro Analysis of Nanoparticle Uptake by Macrophages
Using Chemiluminescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .     255
Sarah L. Skoczen, Timothy M. Potter, and Marina A. Dobrovolskaia
Index . 263