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抗金黄色葡萄球菌感染 scFv-Fc 抗体的制备及其在防治奶牛乳腺炎中的治疗作用。

The preparation and therapeutic roles of scFv-Fc antibody against Staphylococcus aureus infection to control bovine mastitis.

机构信息

Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai JiaoTong University, 800 Dongchuan Road, Shanghai, 200240, China.

出版信息

Appl Microbiol Biotechnol. 2019 Feb;103(4):1703-1712. doi: 10.1007/s00253-018-9548-6. Epub 2019 Jan 3.

Abstract

Staphylococcus aureus-induced bovine mastitis causes significant losses to the dairy industry and available vaccines do not confer adequate protection. As a more attractive alternative, we propose the use of antibody (Ab) therapy. In our previous study, we constructed a bovine single-chain variable fragment (scFv) Ab phage display and successfully obtained scFvs that bound to S. aureus antigens with high affinity. Here, we describe a novel Ab against S. aureus (scFv-Fc Ab). To construct the scFv-Fc Ab, the scFv Ab was genetically fused to the Fc fragment of a bovine IgG1 Ab. Western blot analysis showed that the bovine scFvs-Fc Abs were successfully expressed with horseradish peroxidase-conjugated goat-anti-bovine IgG (Fc) Ab in Escherichia coli cells. The purified bovine scFvs-Fc Abs had good binding activity to S. aureus and effectively inhibited the bacterial growth in culture medium and bovine scFvs-Fc Abs enhanced phagocytosis of S. aureus by neutrophils isolated from peripheral blood in a dose-dependent manner. In the experiment of bovine scFvs-Fc Abs for the treatment of S. aureus-induced bovine mastitis, the total effective percentage reached 82% (9/11). These novel bovine scFvs-Fc Abs may be useful as therapeutic candidates for the prevention and treatment of S. aureus-induced bovine mastitis.

摘要

金黄色葡萄球菌引起的奶牛乳腺炎给奶业造成了巨大损失,而现有的疫苗不能提供充分的保护。作为一种更有吸引力的替代方法,我们建议使用抗体(Ab)治疗。在我们之前的研究中,我们构建了牛单链可变片段(scFv)Ab 噬菌体展示,并成功获得了与金黄色葡萄球菌抗原具有高亲和力结合的 scFvs。在这里,我们描述了一种针对金黄色葡萄球菌的新型 Ab(scFv-Fc Ab)。为了构建 scFv-Fc Ab,scFv Ab 被遗传融合到牛 IgG1 Ab 的 Fc 片段上。Western blot 分析表明,牛 scFvs-Fc Abs 在大肠杆菌细胞中成功地用辣根过氧化物酶标记的山羊抗牛 IgG(Fc)Ab 表达。纯化的牛 scFvs-Fc Abs 对金黄色葡萄球菌具有良好的结合活性,并能有效抑制培养基中细菌的生长,而且牛 scFvs-Fc Abs 能以剂量依赖的方式增强从外周血分离的中性粒细胞对金黄色葡萄球菌的吞噬作用。在牛 scFvs-Fc Abs 治疗金黄色葡萄球菌诱导的奶牛乳腺炎的实验中,总有效率达到 82%(9/11)。这些新型的牛 scFvs-Fc Abs 可能是预防和治疗金黄色葡萄球菌诱导的奶牛乳腺炎的有用治疗候选物。

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