title: Fluorescence polarization-based assay using N-glycan-conjugated quantum dots for screening in hemagglutinin blockers for influenza A viruses.
abstract: Attachment of influenza virus to susceptible cells is mediated by viral protein hemagglutinin (HA), which recognizes cell surface glycoconjugates that terminate in α-sialosides. To develop anti-influenza drugs based on inhibition of HA-mediated infection, novel fluorescent nanoparticles displaying multiple biantennary N-glycan chains with α-sialosides (A2-PC-QDs) that have high affinity for the HA were designed and constructed. The A2-PC-QDs enabled an easy and efficient fluorescence polarization (FP) assay for detection of interaction with the HA and competitive inhibition even by small molecule compounds against A2-PC-QDs-HA binding. The quantum dot (QD)-based FP assay established in the present study is a useful tool for high-throughput screening and to accelerate the development of novel and more effective blockers of the viral attachment of influenza virus.
authors.affiliation: Laboratory of Microbiology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan. okamatsu@vetmed.hokudai.ac.jp
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Okamatsu
authors.forename: Fei
authors.initials: F
authors.lastname: Feng
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Ohyanagi
authors.forename: Noriko
authors.initials: N
authors.lastname: Nagahori
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Someya
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Sakoda
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Miura
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Nishimura
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kida
mesh_terms: D000998:Antiviral Agents / Q000302:isolation & purification* / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D005454:Fluorescence Polarization / Q000379:methods*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000378:metabolism*; D057166:High-Throughput Screening Assays / Q000379:methods; D009980:Influenza A virus / Q000187:drug effects*; D011485:Protein Binding / Q000187:drug effects; D045663:Quantum Dots; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D019267:Hemagglutinin Glycoproteins, Influenza Virus