Switching-peptides for one-step immunoassay and its application to the diagnosis of human hepatitis B

Ji Hong Bong, Hong Rae Kim, Jaeyong Jung, Jun Hee Park, Jeong Soo Sung, Chang Kyu Lee, Kyung Hak Choi, Seong Shick Shin, Min Jung Kang, Hyun Ok Kim, Do Young Lee, Jae Chul Pyun

Research output: Contribution to journalArticlepeer-review

Abstract

Herein, we present switching-peptides for a one-step immunoassay, without the need for additional antibody treatment or washing steps to detect antigen–antibody interactions. Fluorescently labeled switching-peptides were dissociated from the immobilized antibody soon after the antigens were bound to the binding pockets. In this study, four different parts of the antibody (IgG) frame regions were chemically synthesized, and these peptides were bound to immobilized antibodies as switching-peptides. We presented the design principle of switching-peptides and used Pymol software, based on the changes in thermodynamic parameters, to study the interaction between antibodies and switching-peptides. The binding properties of switching-peptides were analyzed based on Förster resonance energy transfer between switching-peptides as well as between switching-peptides and antibodies (IgGs) isolated from different animals. The binding constants of the four switching-peptides to antibodies were estimated to be in the range of 1.48–3.29 μM. Finally, the feasibility of using switching-peptides for the quantitative one-step immunoassay was demonstrated by human hepatitis B surface antigen (hHBsAg) detection and statistical comparison of the assay results with those of conventional ELISA. The limit of detection for HBsAg was determined to be 56 ng/mL, and the dynamic range was estimated to be 136 ng/mL–33 μg/mL. These results demonstrate the feasibility of the one-step immunoassay for HBsAg.

Original languageEnglish
Article number112996
JournalBiosensors and Bioelectronics
Volume178
DOIs
Publication statusPublished - 2021 Apr 15

Bibliographical note

Funding Information:
This work was supported by the National Research Foundation of Korea [grant number: NRF- 2020R1A2B5B01002187 , NRF- 2020R1A5A101913111 ].

Publisher Copyright:
© 2021 Elsevier B.V.

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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