title: High-throughput and sensitive assay for amphotericin B interaction with lipid membrane on the model membrane systems by surface plasmon resonance.
abstract: In the present study, we developed a high-throughput and sensitive assay for interactions of amphotericin B (AmB) with two model lipid membranes, which mimicked mammal cell membrane and fungal membrane using surface plasmon resonance (SPR). The binding kinetics of AmB to the membrane could be analyzed by multiple sensorgrams obtained at different AmB concentrations, indicating that the binding properties could be clarified for an approximately 7-fold concentration range. AmB showed an approximately 18-fold higher affinity for ergosterol-containing membrane than for cholesterol-containing membrane. We also optimized the procedure for the reproducible immobilization of liposome containing the sterols and the estimation of binding kinetics of AmB to the lipid membranes, and the sensitivity of AmB to membrane interaction was 20-fold higher, compared with the reported method. The throughput of the established method for the binding kinetics characterization was calculated to be 10 compounds a day. The results demonstrate that the established SPR method could be a valuable tool for predicting selective binding to sterol-containing membranes.
authors.affiliation: Pharmaceutical Research Center, Shionogi & Co., Ltd., 3-1-1 Futaba-cho, Toyonaka, Osaka 561-0825, Japan. masako.oka@shionogi.co.jp
authors.forename: Masako
authors.initials: M
authors.lastname: Onishi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kamimori
mesh_terms: D000666:Amphotericin B / Q000737:chemistry*; D000935:Antifungal Agents / Q000737:chemistry*; D002784:Cholesterol / Q000737:chemistry*; D004875:Ergosterol / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry; D008563:Membrane Lipids / Q000737:chemistry*; D020349:Surface Plasmon Resonance
chemical_list: D000935:Antifungal Agents; D008081:Liposomes; D008563:Membrane Lipids; D000666:Amphotericin B; D002784:Cholesterol; D004875:Ergosterol