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.