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

Analysis of Complement Activation by Nanoparticles.

Author information

1
Cancer Research Technology Program, Nanotechnology Characterization Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, P.O. Box B, Frederick, MD, 21702, USA.
2
Cancer Research Technology Program, Nanotechnology Characterization Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, P.O. Box B, Frederick, MD, 21702, USA. marina@mail.nih.gov.

Abstract

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

PMID:
29039100
DOI:
10.1007/978-1-4939-7352-1_13
[Indexed for MEDLINE]

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