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1560365 
Journal Article 
A sensitive high throughput ELISA for human eosinophil peroxidase: A specific assay to quantify eosinophil degranulation from patient-derived sources 
Ochkur, SI; Kim, JD; Protheroe, CA; Colbert, D; Condjella, RM; Bersoux, S; Helmers, RA; Moqbel, R; Lacy, P; Kelly, EA; Jarjour, NN; Kern, R; Peters, A; Schleimer, RP; Furuta, GT; Nair, P; Lee, JJ; Lee, NA 
2012 
Yes 
Journal of Immunological Methods
ISSN: 0022-1759 
384 
1-2 
10-20 
22750539 
WOS:000309572300002 
Quantitative high throughput assays of eosinophil-mediated
activities in fluid samples from patients in a clinical setting have been limited to ELISA
assessments for the presence of the prominent granule ribonudeases, ECP and EDN. However, the
demonstration that these ribonudeases are expressed by leukocytes other than eosinophils, as well
as cells of non-hematopoietic origin, limits the usefulness of these assays. Two novel monoclonal
antibodies recognizing eosinophil peroxidase (EPX) were used to develop an eosinophil-specific
and sensitive sandwich ELISA The sensitivity of this EPX-based ELISA was shown to be similar to
that of the commercially available ELISA kits for ECP and EDN. More importantly, evidence is also
presented confirming that among these granule protein detection options, EPX-based ELISA is the
only eosinophil-specific assay. The utility of this high throughput assay to detect released EPX
was shown in ex vivo degranulation studies with isolated human eosinophils. In addition, EPX-
based ELISA was used to detect and quantify eosinophil degranulation in several in vivo patient
settings, including bronchoalveolar lavage fluid obtained following segmental allergen challenge
of subjects with allergic asthma, induced sputum derived from respiratory subjects following
hypotonic saline inhalation, and nasal lavage of chronic rhinosinusitis patients. This unique
EPX-based ELISA thus provides an eosinophil-specific assay that is sensitive, reproducible, and
quantitative. In addition, this assay is adaptable to high throughput formats (e.g, automated
assays utilizing microtiter plates) using the diverse patient fluid samples typically available
in research and clinical settings. (C) 2012 Elsevier B.V. All rights reserved. 
EPX; Eosinophilia; Granule proteins; Allergic inflammation