Characterization of a novel format scFv×VHH single-chain biparatopic antibody against metal binding protein MtsA.

IF 4.5 3区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Protein Science Pub Date : 2024-06-01 DOI:10.1002/pro.5017
Risa Asano, Miyu Takeuchi, Makoto Nakakido, Sho Ito, Chihiro Aikawa, Takeshi Yokoyama, Akinobu Senoo, Go Ueno, Satoru Nagatoishi, Yoshikazu Tanaka, Ichiro Nakagawa, Kouhei Tsumoto
{"title":"Characterization of a novel format scFv×VHH single-chain biparatopic antibody against metal binding protein MtsA.","authors":"Risa Asano, Miyu Takeuchi, Makoto Nakakido, Sho Ito, Chihiro Aikawa, Takeshi Yokoyama, Akinobu Senoo, Go Ueno, Satoru Nagatoishi, Yoshikazu Tanaka, Ichiro Nakagawa, Kouhei Tsumoto","doi":"10.1002/pro.5017","DOIUrl":null,"url":null,"abstract":"<p><p>Biparatopic antibodies (bpAbs) are engineered antibodies that bind to multiple different epitopes within the same antigens. bpAbs comprise diverse formats, including fragment-based formats, and choosing the appropriate molecular format for a desired function against a target molecule is a challenging task. Moreover, optimizing the design of constructs requires selecting appropriate antibody modalities and adjusting linker length for individual bpAbs. Therefore, it is crucial to understand the characteristics of bpAbs at the molecular level. In this study, we first obtained single-chain variable fragments and camelid heavy-chain variable domains targeting distinct epitopes of the metal binding protein MtsA and then developed a novel format single-chain bpAb connecting these fragment antibodies with various linkers. The physicochemical properties, binding activities, complex formation states with antigen, and functions of the bpAb were analyzed using multiple approaches. Notably, we found that the assembly state of the complexes was controlled by a linker and that longer linkers tended to form more compact complexes. These observations provide detailed molecular information that should be considered in the design of bpAbs.</p>","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":null,"pages":null},"PeriodicalIF":4.5000,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11094767/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Protein Science","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1002/pro.5017","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Biparatopic antibodies (bpAbs) are engineered antibodies that bind to multiple different epitopes within the same antigens. bpAbs comprise diverse formats, including fragment-based formats, and choosing the appropriate molecular format for a desired function against a target molecule is a challenging task. Moreover, optimizing the design of constructs requires selecting appropriate antibody modalities and adjusting linker length for individual bpAbs. Therefore, it is crucial to understand the characteristics of bpAbs at the molecular level. In this study, we first obtained single-chain variable fragments and camelid heavy-chain variable domains targeting distinct epitopes of the metal binding protein MtsA and then developed a novel format single-chain bpAb connecting these fragment antibodies with various linkers. The physicochemical properties, binding activities, complex formation states with antigen, and functions of the bpAb were analyzed using multiple approaches. Notably, we found that the assembly state of the complexes was controlled by a linker and that longer linkers tended to form more compact complexes. These observations provide detailed molecular information that should be considered in the design of bpAbs.

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
针对金属结合蛋白 MtsA 的新型格式 scFv×VHH 单链双抗体的表征。
双特异性抗体(bpAbs)是一种工程化抗体,能与同一抗原中的多个不同表位结合。bpAbs 有多种形式,包括基于片段的形式,而针对目标分子的所需功能选择适当的分子形式是一项具有挑战性的任务。此外,优化构建体的设计需要选择适当的抗体模式并调整单个 bpAbs 的连接长度。因此,了解 bpAbs 在分子水平上的特性至关重要。在这项研究中,我们首先获得了针对金属结合蛋白MtsA不同表位的单链可变片段和驼科动物重链可变结构域,然后开发了一种新型格式的单链bpAb,将这些片段抗体与不同的连接体连接起来。我们采用多种方法分析了 bpAb 的理化性质、结合活性、与抗原形成复合物的状态以及功能。值得注意的是,我们发现复合物的组装状态受连接体控制,较长的连接体倾向于形成更紧凑的复合物。这些观察结果提供了详细的分子信息,在设计 bpAb 时应加以考虑。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
Protein Science
Protein Science 生物-生化与分子生物学
CiteScore
12.40
自引率
1.20%
发文量
246
审稿时长
1 months
期刊介绍: Protein Science, the flagship journal of The Protein Society, is a publication that focuses on advancing fundamental knowledge in the field of protein molecules. The journal welcomes original reports and review articles that contribute to our understanding of protein function, structure, folding, design, and evolution. Additionally, Protein Science encourages papers that explore the applications of protein science in various areas such as therapeutics, protein-based biomaterials, bionanotechnology, synthetic biology, and bioelectronics. The journal accepts manuscript submissions in any suitable format for review, with the requirement of converting the manuscript to journal-style format only upon acceptance for publication. Protein Science is indexed and abstracted in numerous databases, including the Agricultural & Environmental Science Database (ProQuest), Biological Science Database (ProQuest), CAS: Chemical Abstracts Service (ACS), Embase (Elsevier), Health & Medical Collection (ProQuest), Health Research Premium Collection (ProQuest), Materials Science & Engineering Database (ProQuest), MEDLINE/PubMed (NLM), Natural Science Collection (ProQuest), and SciTech Premium Collection (ProQuest).
期刊最新文献
Biophysical characterization of RelA-p52 NF-κB dimer-A link between the canonical and the non-canonical NF-κB pathway. Correction to "Award Winners and Abstracts of the 30th Anniversary Symposium of the Protein Society, Baltimore, MD, July 16-19, 2016". On the humanization of VHHs: Prospective case studies, experimental and computational characterization of structural determinants for functionality. A coarse-grained model for disordered and multi-domain proteins. dUTP pyrophosphatases from hyperthermophilic eubacterium and archaeon: Structural and functional examinations on the suitability for PCR application.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1