title: Application of a hemolysis assay for analysis of complement activation by perfluorocarbon nanoparticles.
abstract: Nanoparticles offer new options for medical diagnosis and therapeutics with their capacity to specifically target cells and tissues with imaging agents and/or drug payloads. The unique physical aspects of nanoparticles present new challenges for this promising technology. Studies indicate that nanoparticles often elicit moderate to severe complement activation. Using human in vitro assays that corroborated the mouse in vivo results we previously presented mechanistic studies that define the pathway and key components involved in modulating complement interactions with several gadolinium-functionalized perfluorocarbon nanoparticles (PFOB). Here we employ a modified in vitro hemolysis-based assay developed in conjunction with the mouse in vivo model to broaden our analysis to include PFOBs of varying size, charge and surface chemistry and examine the variations in nanoparticle-mediated complement activity between individuals. This approach may provide the tools for an in-depth structure-activity relationship study that will guide the eventual development of biocompatible nanoparticles.
  
  
  BACKGROUND
  Unique physical aspects of nanoparticles may lead to moderate to severe complement activation in vivo, which represents a challenge to clinical applicability. In order to guide the eventual development of biocompatible nanoparticles, this team of authors report a modified in vitro hemolysis-based assay developed in conjunction with their previously presented mouse model to enable in-depth structure-activity relationship studies.
authors.affiliation: Department of Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Christine T N
authors.initials: CT
authors.lastname: Pham
authors.affiliation: Department of Knowledge, Discovery and Informatics, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Dennis G
authors.initials: DG
authors.lastname: Thomas
authors.affiliation: Department of Ophthalmology and Visual Sciences, Division of Biostatistics, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Julia
authors.initials: J
authors.lastname: Beiser
authors.affiliation: Department of Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Lynne M
authors.initials: LM
authors.lastname: Mitchell
authors.affiliation: Department of Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Huang
authors.affiliation: Department of Medicine, Division of Cardiology, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Angana
authors.initials: A
authors.lastname: Senpan
authors.affiliation: Department of Ophthalmology and Visual Sciences, Division of Biostatistics, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Grace
authors.initials: G
authors.lastname: Hu
authors.affiliation: Department of Ophthalmology and Visual Sciences, Division of Biostatistics, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Mae
authors.initials: M
authors.lastname: Gordon
authors.affiliation: Department of Knowledge, Discovery and Informatics, Pacific Northwest National Laboratory, Richland, WA, USA.
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Baker
authors.affiliation: Department of Medicine, Division of Cardiology, Washington University School of Medicine, St. Louis, MO, USA; Department of Bioengineering, University of Illinois at Urbana-Champaign, Champaign, IL, USA.
authors.forename: Dipanjan
authors.initials: D
authors.lastname: Pan
authors.affiliation: Department of Medicine, Division of Cardiology, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Lanza
authors.affiliation: Department of Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, MO, USA. Electronic address: dhourcad@dom.wustl.edu.
authors.forename: Dennis E
authors.initials: DE
authors.lastname: Hourcade
mesh_terms: D000818:Animals; D003167:Complement Activation / Q000187:drug effects*; D005466:Fluorocarbons / Q000737:chemistry / Q000276:immunology*; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D053758:Nanoparticles / Q000737:chemistry / Q000378:metabolism*; D010316:Particle Size
chemical_list: D005466:Fluorocarbons
keywords: Complement; Hemolysis Assay; Mouse Model; Nanomedicine; Nanoparticles; Perfluorocarbon