Analysis of Complement Activation by Nanoparticles

Methods Mol Biol. 2018;1682:149-160. doi: 10.1007/978-1-4939-7352-1_13.


The complement system is a group of proteins, which function in plasma to assist the innate immunity in rapid clearance of pathogens. The complement system also contributes to coordination of the adaptive immune response. Complement Activation Related Pseudo Allergy or CARPA is a life-threatening condition commonly reported with certain types of drugs and nanotechnology-based combination products. While CARPA symptoms are similar to that of anaphylaxis, the mechanism behind this pathology does not involve IgE and is mediated by the complement system. In vitro assays using serum or plasma derived from healthy donor volunteers correlate with the in vivo complement-mediated reactions, and therefore are helpful in understanding the propensity of a given drug formulation to cause CARPA in patients. In the first edition of this book, we have described an in vitro method for qualitative assessment of the complement activation by nanomaterials using western blotting. Herein, we present a similar method utilizing enzyme-linked immunoassay for quantitative analysis of the complement activation, and we compare the performance of this approach to that of the qualitative western blotting technique. The revised chapter also includes new details about nanoparticle sample preparation.

Keywords: Anaphylaxis; C3; Complement; EIA; Immunoassay; Nanoparticles; Western blot.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Blood Specimen Collection / methods
  • Blotting, Western / methods
  • Complement Activation*
  • Complement C3 / analysis
  • Complement C3 / immunology
  • Humans
  • Immunoenzyme Techniques / methods*
  • Nanoparticles / adverse effects*


  • Complement C3