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(Journal of Leukocyte Biology. 2001;69:1027-1035.)
© 2001 by Society for Leukocyte Biology

Identification and characterization of human eosinophil cationic protein by an epitope-specific antibody

Ester Boix, Esther Carreras, Zoran Nikolovski, Claudi M. Cuchillo and M. Victòria Nogués

Departament de Bioquímica i Biologia Molecular, Facultat de Ciències, Universitat Autònoma de Barcelona, 08193-Bellaterra, Spain

Correspondence: M. Victòria Nogués, Departament de Bioquímica i Biologia Molecular, Facultat de Ciències, Universitat Autònoma de Barcelona, 08193-Bellaterra, Spain. E-mail: victoria.nogues{at}uab.es

The eosinophil cationic protein (ECP) is a basic secretion protein involved in the immune response system. ECP levels in biological fluids are an indicator of eosinophil-specific activation and degranulation and are currently used for the clinical monitoring and diagnosis of inflammatory disorders. A polyclonal epitope-specific antibody has been obtained by immunizing rabbits with a conjugated synthetic peptide. A sequence corresponding to a large exposed loop in the human ECP three-dimensional structure (D115–Y122) was selected as a putative antigenic epitope. The antibody was purified on an affinity column using recombinant ECP (rECP) as antigen. The antibody (D112–P123 Ab) specifically recognizes rECP and its native glycosylated and nonglycosylated forms in plasma, granulocytes, and sputum. The antibody detects as little as 1 ng of rECP, can be used both in reducing and nonreducing conditions, and does not cross-react with the highly homologous eosinophil-derived neurotoxin or other proteins of the pancreatic ribonuclease superfamily.

Key Words: immunoassay • granulocytes • plasma • sputum




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