Quantification of Porcine Complement Activation Fragment C3a by a Neoepitope-Based Enzyme-Linked Immunosorbent Assay

Per H Nilsson; Kristin Pettersen; Martin Oppermann; Espen W Skjeflo; Hilde Fure; Dorte Christiansen; Tom Eirik Mollnes

doi:10.1007/978-1-0716-1016-9_5

Quantification of Porcine Complement Activation Fragment C3a by a Neoepitope-Based Enzyme-Linked Immunosorbent Assay

Methods Mol Biol. 2021:2227:51-59. doi: 10.1007/978-1-0716-1016-9_5.

Authors

Per H Nilsson^{1

2}, Kristin Pettersen³, Martin Oppermann⁴, Espen W Skjeflo^{3

5}, Hilde Fure³, Dorte Christiansen³, Tom Eirik Mollnes^{6

5

7}

Affiliations

¹ Department of Immunology, University of Oslo and Oslo University Hospital Rikshospitalet, Oslo, Norway. per.nilsson@medisin.uio.no.
² Linnaeus Centre for Biomaterials Chemistry, Linnaeus University, Kalmar, Sweden. per.nilsson@medisin.uio.no.
³ Research Laboratory, Nordland Hospital, Bodø, Norway.
⁴ Institute for Cellular and Molecular Immunology, University Medical Center, Georg-August-University Göttingen, Göttingen, Germany.
⁵ Faculty of Health Sciences, K.G Jebsen TREC, UiT-The Arctic University of Norway, Tromsø, Norway.
⁶ Department of Immunology, University of Oslo and Oslo University Hospital Rikshospitalet, Oslo, Norway.
⁷ Centre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, Norway.

PMID: 33847930
DOI: 10.1007/978-1-0716-1016-9_5

Abstract

Enzyme-linked immunosorbent assay (ELISA) enables fast and simple quantification of analytes in the pico- to nanogram range in complex samples. Here, we describe an ELISA for the detection of porcine C3a as a marker for complement activation. Antibody specificity is critical for a robust assay. This assay is based on a pair of antibodies specific for the porcine C3a molecule and thus does not react with native C3.

Keywords: Anaphylatoxin; C3a; Complement; Complement activation; ELISA; Porcine.

MeSH terms

Animals
Antibodies, Monoclonal / metabolism
Antibody Specificity
Complement Activation / physiology
Complement C3a / analysis*
Complement C3a / metabolism
Cross Reactions / immunology
Enzyme-Linked Immunosorbent Assay / methods
Epitopes / metabolism
Goats
Mice
Sepsis / blood
Sepsis / diagnosis
Sepsis / veterinary
Swine / blood*
Swine / immunology
Swine Diseases / blood
Swine Diseases / diagnosis

Substances

Antibodies, Monoclonal
Epitopes
Complement C3a