Bong, Ji-Hong Kim, Hong-Rae Jung, Jaeyong Park, Jun-Hee Sung, Jeong Soo Lee, Chang Kyu Choi, Kyung-Hak Shin, Seong-Shick Kang, Min-Jung Kim, Hyun Ok Lee, Do Young Pyun, Jae-Chul 2024-01-19T15:01:28Z 2024-01-19T15:01:28Z 2021-09-05 2021-04-15 0956-5663 https://pubs.kist.re.kr/handle/201004/117126 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 Forster 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 mu M. 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 mu g/mL. These results demonstrate the feasibility of the one-step immunoassay for HBsAg. English ELSEVIER ADVANCED TECHNOLOGY Switching-peptides for one-step immunoassay and its application to the diagnosis of human hepatitis B Article 10.1016/j.bios.2021.112996 1 BIOSENSORS & BIOELECTRONICS, v.178 BIOSENSORS & BIOELECTRONICS 178 scie scopus 000621206100005 2-s2.0-85099960149 Biophysics Biotechnology & Applied Microbiology Chemistry, Analytical Electrochemistry Nanoscience & Nanotechnology Biophysics Biotechnology & Applied Microbiology Chemistry Electrochemistry Science & Technology - Other Topics Article Switching-peptide One-step immunoassay Frame region Hepatitis Medical diagnosis