Human eosinophil peroxidase: purification and characterization
1985 (English)In: Journal of Immunology, ISSN 0022-1767, E-ISSN 1550-6606, Vol. 134, no 3, 1875-1879 p.Article in journal (Refereed) Published
Human eosinophil peroxidase (EPO) was isolated from granules from granulocytes of a patient with hypereosinophilia. The granules were extracted by means of 0.2 M NaAc, pH 4.0. The purification steps included gel filtration chromatography on Sephadex G-75 superfine and ion-exchange chromatography on CM-Sephadex G-50. The purified protein showed one band on agarose-electrophoresis, a high peroxidase activity, and a 415-nm/280 nm ratio of 1.15. After reduction, EPO showed two bands on SDS-PAGE of m.w. 52,000 and 15,000, respectively. On gel filtration, the unreduced protein had a m.w. of approximately 77,000. Amino acid analyses showed a high content of arginine and aspartic acid. Monospecific antibodies to EPO were prepared in rabbits, and a specific radioimmunoassay was developed. There was an almost linear correlation between the content of EPO measured by the radioimmunoassay and the number of eosinophils in a mixed cell extract from reference material, indicating the eosinophil origin of EPO. The content of EPO was estimated to be 15.0 micrograms/10(6) eosinophils.
Place, publisher, year, edition, pages
1985. Vol. 134, no 3, 1875-1879 p.
Amino Acids/analysis, Antigen-Antibody Reactions, Chromatography; Gel, Eosinophil Peroxidase, Eosinophilia/enzymology, Eosinophils/*enzymology, Humans, Immunodiffusion, Leukocyte Count, Molecular Weight, Peroxidases/analysis/*isolation & purification/metabolism, Radioimmunoassay, Research Support; Non-U.S. Gov't
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:uu:diva-74328PubMedID: 3918110OAI: oai:DiVA.org:uu-74328DiVA: diva2:102238