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. 2022;16(3):334-341.
doi: 10.1007/s13206-022-00076-x. Epub 2022 Jul 27.

One-Step Homogeneous Immunoassay for the Detection of Influenza Virus Using Switching Peptide and Graphene Quencher

Hong-Rae Kim #  1   2 , Ji-Hong Bong #  1 , Tae-Hun Kim  1 , Seung-Shick Shin  3 , Min-Jung Kang  4 , Won-Bo Shim  5 , Do Young Lee  3 , Dong Hee Son  2 , Jae-Chul Pyun  1
Affiliations

One-Step Homogeneous Immunoassay for the Detection of Influenza Virus Using Switching Peptide and Graphene Quencher

Hong-Rae Kim et al. Biochip J. 2022.

Abstract

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.

Keywords: Influenza-A and influenza-B virus; Lateral flow immunoassay; One-step homogeneous immunoassay; Switching peptide.

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Conflict of interest statement

Conflict of interestsThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
One-step immunoassay based on switching peptide. a Switching peptide. b Configuration of one-step immunoassay based on switching peptide
Fig. 2
Fig. 2
Optimization of assay conditions. a Fluorescence quenching with graphene. b Loading concentration of switching peptide. c pH stability. d Stability in salt. Selectivity of antibodies against e Inf-A and Inf-B.
Fig. 3
Fig. 3
One-step immunoassay of a Influenza-A and b Influenza-B in PBS and serum
Fig. 4
Fig. 4
Comparison of one-step immunoassay of a Inf-A and b Inf-B with lateral flow immunoassay
See this image and copyright information in PMC

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