An inhibition enzyme immunoassay for quantification of blood group antigen in stroma-free hemoglobin

A. B.E. Hideki, Kunihiko Nakai, Tsuneo A. Takahashi, Sadayoshi Sekiguchi

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1 Citation (Scopus)


A sensitive enzyme immunoassay (EIA) for quantifying blood group antigen was developed for the quality control of stroma-free hemoglobin (SF-Hb). Glycophorin A was purified from type A red blood cells and used as the coating antigen and the standard for EIA. The lower detection limit was 0.3 ng of purified glycophorin A - a value which was estimated to be 10-fold more sensitive than that by the hemagglutination inhibition (HAI) test. This EIA was applied to evaluate six representative SF-Hb solutions: conventional SF-Hb prepared by filtration with a 0.22 μm filter and five other SF-Hbs prepared by an additional technique including either heating, acid precipitation, chloroform extraction, ultrafiltration or filtration with a virus-removal filter. Conventional SF-Hb and the heat-treated SF-Hb contained a significant amount of antigen at 8.40-8.70 ng/mg of hemoglobin and were also positive by the HAI test. In contrast,The other four SF-Hb were negative by the HAI test, but EIA successfully revealed that they still contained the antigen in the range of 0.46-0.84 ng/mg and that their purity was more than 99.98%. These results indicate that these SF-Hb are 10 times purer than conventional SF-Hb. EIA is sensitive, fast and easy to perform, and is therefore a useful tool for the quality control of SF-Hb.

Original languageEnglish
Pages (from-to)19-28
Number of pages10
JournalArtificial Organs Today (Japan)
Issue number1
Publication statusPublished - 1996 Dec 1
Externally publishedYes


  • Blood group antigen
  • Inhibition enzyme immunoassay
  • Stroma-free hemoglobin

ASJC Scopus subject areas

  • Biomedical Engineering


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