IF 3.2 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Wei-Chu Wang, Chi-Hsin Lee, Chao-Jung Wu, Sy-Jye Leu, Pei-Shih Kao, Bor-Yu Tsai, Ko-Jiunn Liu, Yu-Wei Chiang, Hsiu-Jung Lo, Yan-Chiao Mao, Yi-Yuan Yang
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引用次数: 0

摘要

金黄色葡萄球菌是一种在人类中普遍存在的革兰氏阳性细菌,具有导致严重机会性感染和抗生素耐药性增加的巨大风险。金黄色葡萄球菌中的α-烯醇化酶在细胞外基质结合和生物膜形成中发挥着重要作用。这些功能使金黄色葡萄球菌能够侵入宿主组织并导致感染。本研究旨在通过噬菌体展示技术开发针对金黄色葡萄球菌表面蛋白的特异性α-烯醇化酶鸡抗体,作为抗生素治疗的潜在替代品。用重组金黄色葡萄球菌α-烯醇化酶对鸡进行免疫,从而构建了两个噬菌体展示单链可变片段库,分别包含 3.32 × 106 和 8.60 × 105 个不同连接子长度的转化子。经过四轮生物筛选,选出了五个单链可变片段抗体克隆,其中包括三个具有高结合亲和力的克隆(SaS1、SaS2 和 SaL2)。这些克隆在表位图谱和交叉反应试验中表现出不同的结合模式,其中 SaS1 和 SaS2 能特异性识别金黄色葡萄球菌α-烯醇化酶,SaL2 能与肺炎链球菌α-烯醇化酶发生交叉反应。此外,还通过细胞酶联免疫吸附试验(ELISA)和流式细胞术试验验证了这些抗体克隆对临床金黄色葡萄球菌菌株(包括甲氧西林敏感菌株和甲氧西林耐药菌株)的特异性。SaS1、SaS2 和 SaL2 的鉴定强调了它们的诊断和治疗潜力,为传统抗生素疗法提供了有前途的替代品。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Phage Display Selected Chicken Antibodies Targeting Surface Alpha Enolase in Staphylococcus aureus

Staphylococcus aureus, a prevalent gram-positive bacterium in human populations, poses a significant risk for causing serious opportunistic infections and increasing antibiotic resistance. Alpha-enolase in S. aureus plays important roles in extracellular matrix binding and biofilm formation. These functions enable S. aureus to invade host tissues and cause infections. The aim of this study was to develop specific alpha-enolase chicken antibodies through phage display technology targeting S. aureus surface proteins as a potential alternative to antibiotic therapy. A chicken was immunized with recombinant S. aureus alpha-enolase, leading to the construction of two phage display single-chain variable fragment libraries of 3.32 × 106 and 8.60×105 transformants with different linker lengths. After four rounds of biopanning, five single-chain variable fragment antibody clones, including three with high binding affinities (SaS1, SaS2, and SaL2), were selected. These clones exhibited distinct binding patterns in epitope mapping and cross-reaction assays, with SaS1 and SaS2 specifically recognizing S. aureus alpha-enolase and SaL2 cross-reacting with Streptococcus pneumoniae alpha-enolase. Furthermore, the specificity of these antibody clones toward clinical S. aureus strains, including methicillin-sensitive and methicillin-resistant strains, was validated through cell-based enzyme-linked immunosorbent assays (ELISA) and flow cytometry assays. The identification of SaS1, SaS2, and SaL2 underscores their diagnostic and therapeutic potential, offering promising alternatives to traditional antibiotic therapies.

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来源期刊
Biotechnology Journal
Biotechnology Journal Biochemistry, Genetics and Molecular Biology-Molecular Medicine
CiteScore
8.90
自引率
2.10%
发文量
123
审稿时长
1.5 months
期刊介绍: Biotechnology Journal (2019 Journal Citation Reports: 3.543) is fully comprehensive in its scope and publishes strictly peer-reviewed papers covering novel aspects and methods in all areas of biotechnology. Some issues are devoted to a special topic, providing the latest information on the most crucial areas of research and technological advances. In addition to these special issues, the journal welcomes unsolicited submissions for primary research articles, such as Research Articles, Rapid Communications and Biotech Methods. BTJ also welcomes proposals of Review Articles - please send in a brief outline of the article and the senior author''s CV to the editorial office. BTJ promotes a special emphasis on: Systems Biotechnology Synthetic Biology and Metabolic Engineering Nanobiotechnology and Biomaterials Tissue engineering, Regenerative Medicine and Stem cells Gene Editing, Gene therapy and Immunotherapy Omics technologies Industrial Biotechnology, Biopharmaceuticals and Biocatalysis Bioprocess engineering and Downstream processing Plant Biotechnology Biosafety, Biotech Ethics, Science Communication Methods and Advances.
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