A sensitive sandwich ELISA for measuring thrombopoietin in human serum: Serum thrombopoietin levels in healthy volunteers and in patients with haemopoietic disorders

Tomoyuki Tahara, Kensuke Usuki, Hiroaki Sato, Hideya Ohashi, Haruhiko Morita, Haruhiko Tsumura, Atsushi Matsumoto, Hiroshi Miyazaki, Akio Urabe, Takashi Kato

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188 Citations (Scopus)

Abstract

A sensitive sandwich enzyme-linked immunosorbent assay (ELISA) has been established to estimate serum thrombopoietin (TPO) concentrations in healthy volunteers and patients with haemopoietic disorders. The ELISA uses a mouse monoclonal antibody (Ab) as the capture Ab and a biotinylated rabbit polyclonal Ab as the detector. The ELISA was reproducible, highly sensitive and specific for human TPO. The coefficients of intra- and interassay variation were from 3.0% to 4.9% and from 5.9% to 6.1%, respectively. The quantitative limit of the ELISA was 0.09 fmol/ml in serum. The quantitative limit was lower than the normal level. The dose-response curves of serum samples from healthy volunteers and patients with haemopoietic disorders were parallel to the standard curves. The ELISA did not cross-react with a variety of blood components and cytokines to produce false-positive results. The serum TPO concentrations from 29 normal males and 21 females were 0.79 ± 0.35 and 0.70 ± 0.26 fmol/ml, respectively. Serum TPO levels in patients with aplastic anaemia (AA), acute lymphocytic leukaemia (ALL) and essential thrombocythaemia (ET) were measured using the ELISA. The serum TPO levels in the patients with ET (n = 6, 2.80 ± 1.55 fmol/ml) were higher than the normal level. The patients with AA (n = 7, 18.53 ± 12.37 fmol/ml) and ALL (n = 5, 10.36 ± 5.57 fmol/ml) had significantly higher serum TPO levels than normal individuals. These results indicate that the ELISA specific to TPO should prove useful in measuring the TPO concentration in serum samples.

Original languageEnglish
Pages (from-to)783-788
Number of pages6
JournalBritish Journal of Haematology
Volume93
Issue number4
DOIs
Publication statusPublished - 1996 Jan 1

Keywords

  • ELISA
  • c-Mpl ligand
  • haemopoietic disorders
  • thrombopoietin

ASJC Scopus subject areas

  • Hematology

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