Production of monoclonal antibodies against human erythropoietin and their use in the purification of human urinary erythropoietin

Miyazaki Hiroshi Miyazaki, Kozutsumi Hiroyuki Kozutsumi, Kato Takashi Kato, Hoshi Sakuo Hoshi, Tamura Setsuko Tamura, Kubota Mamoru Kubota, Suzuki Takamoto Suzuki

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Several murine monoclonal antibodies (MAbs) specific for human erythropoietin (HuEpo) were produced by hybridomas obtained from the fusion of murine myeloma cells, P3X63-Ag.8-653, with the splenocytes of mice immunized with recombinant human Epo (rHuEpo). Based on epitope analysis by a competitive binding assay, these MAbs could be classified into at least three groups: (1) 1E10, (2) 1H7, (3) 2D6, 3D6 and 3D8. In a sandwich enzyme-linked immunosorbent assay (ELISA), using these MAbs as the solid-phase antibodies, MAb-bound HuEpo was detected with rabbit anti-HuEpo sera. Some combinations of two different classes of MAbs, such as 1H7 and 3D8, were found to capture much more HuEpo than each MAb used individually. Urinary HuEpo (U-HuEpo) was highly purified from the urine of patients with severe aplastic anemia with about 50% final recovery using an immunoaffinity column on which a mixture of 1H7 and 3D8 was immobilized. The purified U-HuEpo had a specific activity of 77 340 U/mg in a radioimmunoassay (RIA) and of 76 673 U/mg using an in vivo bioassay.

Original languageEnglish
Pages (from-to)261-267
Number of pages7
JournalJournal of Immunological Methods
Volume113
Issue number2
DOIs
Publication statusPublished - 1988 Oct 26
Externally publishedYes

Keywords

  • Erythropoietin
  • Immunoaffinity chromatography
  • Monoclonal antibody

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Fingerprint Dive into the research topics of 'Production of monoclonal antibodies against human erythropoietin and their use in the purification of human urinary erythropoietin'. Together they form a unique fingerprint.

  • Cite this

    Hiroshi Miyazaki, M., Hiroyuki Kozutsumi, K., Takashi Kato, K., Sakuo Hoshi, H., Setsuko Tamura, T., Mamoru Kubota, K., & Takamoto Suzuki, S. (1988). Production of monoclonal antibodies against human erythropoietin and their use in the purification of human urinary erythropoietin. Journal of Immunological Methods, 113(2), 261-267. https://doi.org/10.1016/0022-1759(88)90340-7