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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 
 BIOSENSORS & BIOELECTRONICS, Vol.178 : 112996, 2021-04 
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Frame region ; Hepatitis ; Medical diagnosis ; One-step immunoassay ; Switching-peptide
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.
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1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
Yonsei Authors
Kim, Hyun Ok(김현옥) ORCID logo https://orcid.org/0000-0002-4964-1963
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