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

研究成果: Article

10 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)261-267
ページ数7
ジャーナルJournal of Immunological Methods
113
2
DOI
出版ステータスPublished - 1988 10 26
外部発表はい

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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