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A monoclonal antibody-based sandwich enzyme-linked immunosorbent assay for detection of secreted alpha-defensin

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Title: A monoclonal antibody-based sandwich enzyme-linked immunosorbent assay for detection of secreted alpha-defensin
Authors: Nakamura, Kiminori Browse this author →KAKEN DB
Sakuragi, Naoya Browse this author
Ayabe, Tokiyoshi Browse this author →KAKEN DB
Keywords: Sandwich ELISA
Monoclonal antibodies
Paneth cell
Inflammatory bowel disease
Issue Date: 15-Dec-2013
Publisher: Academic press inc elsevier science
Journal Title: Analytical biochemistry
Volume: 443
Issue: 2
Start Page: 124
End Page: 131
Publisher DOI: 10.1016/j.ab.2013.08.021
PMID: 23994564
Abstract: Paneth cells at the base of small intestinal crypts secrete alpha-defensins, which contribute to innate immunity and shape composition of enteric microbiota. Efforts to establish a relationship between secreted alpha-defensins and disease have been hampered by a lack of sensitive assays to quantify luminal alpha-defensins. Here we report on a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) for the mouse Paneth cell alpha-defensin cryptdin-4 (Crp4) in varied sources, including luminal contents rinsed from stomach to distal colon and fecal pellets. One pair of monoclonal antibodies (mAbs), selected from 10 rat hybridomas secreting Crp4-specific mAbs, was optimized for Crp4 detection and specificity in the sandwich ELISA. In CD1 mice, luminal Crp4 levels increased gradually from 6.8 +/- 5.2 ng/ml in proximal small intestine to 54.3 +/- 10.3 ng/ml in distal small intestine, and the peptide was detected in colonic lumen and feces. Secreted Crp4 was reduced significantly in feces of IL10 null mice, a model of inflammatory bowel disease (IBD) when compared with wild-type controls. This Crp4 sandwich ELISA enables accurate determinations of luminal alpha-defensins as biomarkers of Paneth cell function and enteric integrity in diverse disease states such as IBD, infectious disease, graft versus host disease, and obesity in association with dysbiosis of the intestinal microbiota. (C) 2013 Elsevier Inc. All rights reserved.
Type: article (author version)
Appears in Collections:生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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