Improved enzyme-linked immunosorbent assay using surface-adhesive antibody-oriented immobilizing biolinker: a proof-of-concept study

IF 2.5 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Ae Sol Lee, Hye Ryoung Heo, Chang Sup Kim, Hyung Joon Cha
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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.

Abstract Image

使用面向抗体的表面粘附固定生物链接剂改进酶联免疫吸附测定法:概念验证研究
酶联免疫吸附试验(ELISA)已被广泛应用于临床和研究实验室检测与疾病相关的抗原。酶联免疫吸附法的主要缺点之一是利用物理吸附将抗体固定在表面上,导致灵敏度、重现性和精确度较低。在本研究中,我们将由抗体固定蛋白 A 的 BC 结构域和表面粘附性贻贝粘附蛋白(MAP)组成的 BC-MAP 连接器应用于 ELISA 平台,以克服这些局限性。使用 BC-MAP 连接物的 ELISA 性能与未经处理的 ELISA 性能进行了比较。与利用抗体物理吸附的未经处理的 ELISA 相比,BC-MAP 蛋白在暴露 BC 结构域的同时被重复涂布到表面,从而使 ELISA 的灵敏度和重复性提高了两倍。因此,所提出的方法可以成功地用于 ELISA 平台,以诊断和管理疾病。
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来源期刊
Biotechnology and Bioprocess Engineering
Biotechnology and Bioprocess Engineering 工程技术-生物工程与应用微生物
CiteScore
5.00
自引率
12.50%
发文量
79
审稿时长
3 months
期刊介绍: Biotechnology and Bioprocess Engineering is an international bimonthly journal published by the Korean Society for Biotechnology and Bioengineering. BBE is devoted to the advancement in science and technology in the wide area of biotechnology, bioengineering, and (bio)medical engineering. This includes but is not limited to applied molecular and cell biology, engineered biocatalysis and biotransformation, metabolic engineering and systems biology, bioseparation and bioprocess engineering, cell culture technology, environmental and food biotechnology, pharmaceutics and biopharmaceutics, biomaterials engineering, nanobiotechnology, and biosensor and bioelectronics.
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