Biotechnology and Bioprocess Engineering 2024; 29(3): 543-550  
Improved enzyme-linked immunosorbent assay using surface-adhesive antibody-oriented immobilizing biolinker: a proof-of-concept study
Ae Sol Lee1 · Hye Ryoung Heo1 · Chang Sup Kim1 · Hyung Joon Cha2,3
1 Department of Chemical and Biochemical Engineering, Dongguk University, Seoul 04620, Republic of Korea
2 Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 37673, Republic of Korea
3 Medical Science and Engineering, School of Convergence Science and Technology, Pohang University of Science and Technology, Pohang 37673, Republic of Korea
Correspondence to: Chang Sup Kim
biocskim@dgu.ac.kr
Hyung Joon Cha
hjcha@postech.ac.kr

Ae Sol Lee and Hye Ryoung Heo have contributed equally to this work.
Received: September 27, 2023; Revised: March 6, 2024; Accepted: March 19, 2024; Published online: March 27, 2024.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Enzyme-linked immunosorbent assays (ELISA) have been widely used to detect disease-related antigens in clinical and research laboratories. One of the main drawbacks of ELISA is the utilization of physical adsorption for immobilizing antibodies on a surface, causing low sensitivity, reproducibility, and precision. In this study, we applied a BC-MAP linker composed of antibody-immobilizing BC domains of protein A and surface-adhesive mussel adhesive protein (MAP) to an ELISA platform to overcome these limitations. The performance of ELISA using BC-MAP linker was compared with that of untreated ELISA. BC-MAP proteins were reproducibly coated to the surface while exposing BC domains, resulting in twofold higher sensitivity and improved reproducibility of ELISA compared to the untreated ELISA utilizing physical adsorption of antibodies. Thus, the proposed method could be successfully used in ELISA platforms to diagnose and manage diseases.
Keywords: Enzyme-linked immunosorbent assays · Orientated antibody immobilization · Surface adhesive antibody-oriented immobilizing linker · Sensitivity · Reproducibility

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