Kim Hong-Rae, Bong Ji-Hong, Kim Tae-Hun, Shin Seung-Shick, Kang Min-Jung, Shim Won-Bo, Lee Do Young, Son Dong Hee, Pyun Jae-Chul
Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-Ro, Seodaemun-gu, Seoul, 03722 Korea.
Department of Chemistry, Texas A&M University, College Station, TX 77843 USA.
Biochip J. 2022;16(3):334-341. doi: 10.1007/s13206-022-00076-x. Epub 2022 Jul 27.
One-step homogeneous immunoassay was developed for detecting influenza viruses A and B (Inf-A and Inf-B) using the switching peptide H2. As the fluorescence-labeled switching peptide dissociated from the binding pocket of detection antibodies, the fluorescence signal could be directly generated by the binding of Inf-A and Inf-B without washing (i.e., one-step immunoassay). For the one-step homogeneous immunoassay with detection antibodies in solution, graphene was labeled with the antibodies as a fluorescence quencher. To test the feasibility of the homogeneous one-step immunoassay, the stability of the antibody complex with the switching peptide was evaluated under different pH and salt conditions. The one-step homogeneous immunoassay with switching peptide was conducted using influenza virus antigens in phosphate-buffered saline and real samples with inactivated Inf-A and Inf-B spiked in serum. Finally, the one-step homogeneous immunoassay results were compared with those of commercially available lateral flow immunoassays.
利用开关肽H2开发了一种用于检测甲型和乙型流感病毒（Inf-A和Inf-B）的一步均相免疫测定法。由于荧光标记的开关肽从检测抗体的结合口袋解离，Inf-A和Inf-B的结合可直接产生荧光信号，无需洗涤（即一步免疫测定法）。对于溶液中检测抗体的一步均相免疫测定，石墨烯用抗体标记作为荧光猝灭剂。为了测试均相一步免疫测定的可行性，在不同的pH和盐条件下评估了抗体与开关肽复合物的稳定性。使用磷酸盐缓冲盐水中的流感病毒抗原以及添加了灭活Inf-A和Inf-B的血清实际样品进行了含开关肽的一步均相免疫测定。最后，将一步均相免疫测定结果与市售侧向流动免疫测定结果进行了比较。
