Novel screening systems for HIV-1 fusion mediated by two extra-virion heptad repeats of gp41

Hiroki Nishikawa, Eiichi Kodama, Ayako Sakakibara, Ayako Fukudome, Kazuki Izumi, Shinya Oishi, Nobutaka Fujii, Masao Matsuoka

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9 Citations (Scopus)


Entry of human immunodeficiency virus type 1 (HIV-1) into target cells is mediated by its envelope protein gp41 through membrane fusion. Interaction of two extra-virion heptad repeats (HRs) in the gp41 plays a pivotal role in the fusion, and its inhibitor, enfuvirtide (T-20), blocks HIV-1 entry. To identify agents that block HIV-1 fusion, two screening methods based on detection and quantification by the enzyme-linked immunosorbent assay (ELISA) principle have been established. One method uses an alkaline phosphatase (ALP)-conjugated antibody (Ab-ELISA) and the other uses an ALP-fused HR (F-ELISA) to detect and quantify the interaction of the two HRs. The F-ELISA was more simple and rapid, since no ALP-conjugated antibody reaction was required. Both ELISAs detected all the fusion inhibitors tested except for T-20. Interaction of the two HRs was observed in both ELISAs, even in the presence of 10% dimethyl sulfoxide. Ab-ELISA performed best in a pH ranging from 6 to 8, while F-ELISA performed best at a pH ranging from 7 to 8. These results indicate that both established ELISAs are suitable for the identification of HIV-1 fusion inhibitors.

Original languageEnglish
Pages (from-to)71-76
Number of pages6
JournalAntiviral Research
Issue number1
Publication statusPublished - 2008 Oct


  • Alkaline phosphatase
  • Fusion
  • Gp41
  • HIV-1
  • Screening


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