Suppr超能文献

基于 VH 抗体片段的乳胶凝集试验检测猪链球菌 2 型的建立。

Development of a latex agglutination test based on VH antibody fragment for detection of Streptococcus suis serotype 2.

机构信息

The Medical Microbiology Program, Graduate School, Chulalongkorn University, Bangkok, Thailand.

Department of Veterinary Public Health, Faculty of Veterinary Sciences, Chulalongkorn University, Bangkok, Thailand.

出版信息

PLoS One. 2024 Apr 3;19(4):e0299691. doi: 10.1371/journal.pone.0299691. eCollection 2024.

Abstract

Streptococcus suis serotype 2 (SS2) is an important porcine pathogen that causes diseases in both swine and human. For rapid SS2 identification, a novel latex agglutination test (LAT) based on heavy-chain variable domain antibody (VH) was developed. Firstly, the soluble 47B3 VH antibody fragment from a phage display library, in which cysteine residues were engineered at the C-terminus, was expressed in Escherichia coli. The purified protein was then gently reduced to form monomeric soluble 47B3 VH subsequently used to coat with latex beads by means of site-specific conjugation. The resulting VH-coated beads gave a good agglutination reaction with SS2. The LAT was able to distinguish S. suis serotype 2 from serotype 1/2, which shares some common sugar residues, and showed no cross-reaction with other serotypes of S. suis or other related bacteria. The detection sensitivity was found to be as high as 1.85x106 cells. The LAT was stable at 4°C for at least six months without loss of activity. To the best of our knowledge, this is the first LAT based on a VH antibody fragment that can be considered as an alternative for conventional antibody-based LAT where VHs are the most favored recombinant antibody.

摘要

猪链球菌血清 2 型(SS2)是一种重要的猪病原体,可引起猪和人类的疾病。为了快速鉴定 SS2,开发了一种基于重链可变区抗体(VH)的新型乳胶凝集试验(LAT)。首先,在大肠杆菌中表达了来自噬菌体展示文库的可溶性 47B3 VH 抗体片段,该文库在 C 末端设计了半胱氨酸残基。然后,将纯化的蛋白质温和还原形成单体可溶性 47B3 VH,随后通过位点特异性缀合将其涂覆在乳胶珠上。所得 VH 涂覆的珠粒与 SS2 产生良好的凝集反应。LAT 能够区分 SS2 血清型 1/2,因为它们具有一些共同的糖残基,并且与其他 SS2 血清型或其他相关细菌没有交叉反应。检测灵敏度高达 1.85x106 个细胞。LAT 在 4°C 下至少稳定六个月,而不会失去活性。据我们所知,这是第一种基于 VH 抗体片段的 LAT,可以替代传统的基于抗体的 LAT,其中 VH 是最受欢迎的重组抗体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9630/10990187/3631d8d2a04e/pone.0299691.g001.jpg

文献检索

告别复杂PubMed语法,用中文像聊天一样搜索,搜遍4000万医学文献。AI智能推荐,让科研检索更轻松。

立即免费搜索

文件翻译

保留排版,准确专业,支持PDF/Word/PPT等文件格式,支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述,25分钟生成高质量综述,智能提取关键信息,辅助科研写作。

立即免费体验