Expression, purification and characterization of anti-FGF7 domain antibody identified using phage display technique

Q4 Pharmacology, Toxicology and Pharmaceutics

Infarma Pharmaceutical Sciences Pub Date : 2021-10-27 DOI:10.34172/ps.2021.64

Aliakbar Alizadeh, Mona Roshani, Omid Jamshidi Kandjani, Milad Soltani-Saif, S. Dastmalchi

{"title":"Expression, purification and characterization of anti-FGF7 domain antibody identified using phage display technique","authors":"Aliakbar Alizadeh, Mona Roshani, Omid Jamshidi Kandjani, Milad Soltani-Saif, S. Dastmalchi","doi":"10.34172/ps.2021.64","DOIUrl":null,"url":null,"abstract":"Background: Fibroblast growth factors (FGFs) are involved in angiogenesis, wound healing and embryonic development. However, one of the causes of cancer cell growth in fibroblast-dependent cancers is FGF7 secreted by fibroblasts. Therefore, antibodies against FGF7 can be used for treatment of these types of cancers. Methods: In previous studies, a phage displaying single domain antibody, D53, against human FGF7 has been identified using the phage display technique. In the present study, D53 was produced and purified in its isolated form. ELISA experiment was performed to evaluate the binding of D53 to FGF7. The mode of interaction of D53-FGF7 was explored using docking study and molecular dynamics (MD) simulations. Results: The expression and purification processes were verified using western blotting and SDS-PAGE analyses. ELISA experiment showed that D53 is able to recognize and bind FGF7. Docking study and MD simulations indicated that compared to dummy VH, D53 has more affinity towards FGF7. Conclusion: The findings in the current study can be useful for generation and development of FGF7 inhibitors with potential use in fibroblast-dependent cancers.","PeriodicalId":31004,"journal":{"name":"Infarma Pharmaceutical Sciences","volume":"6 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2021-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Infarma Pharmaceutical Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.34172/ps.2021.64","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Pharmacology, Toxicology and Pharmaceutics","Score":null,"Total":0}

引用次数: 0

Abstract

Background: Fibroblast growth factors (FGFs) are involved in angiogenesis, wound healing and embryonic development. However, one of the causes of cancer cell growth in fibroblast-dependent cancers is FGF7 secreted by fibroblasts. Therefore, antibodies against FGF7 can be used for treatment of these types of cancers. Methods: In previous studies, a phage displaying single domain antibody, D53, against human FGF7 has been identified using the phage display technique. In the present study, D53 was produced and purified in its isolated form. ELISA experiment was performed to evaluate the binding of D53 to FGF7. The mode of interaction of D53-FGF7 was explored using docking study and molecular dynamics (MD) simulations. Results: The expression and purification processes were verified using western blotting and SDS-PAGE analyses. ELISA experiment showed that D53 is able to recognize and bind FGF7. Docking study and MD simulations indicated that compared to dummy VH, D53 has more affinity towards FGF7. Conclusion: The findings in the current study can be useful for generation and development of FGF7 inhibitors with potential use in fibroblast-dependent cancers.

查看原文本刊更多论文

利用噬菌体展示技术鉴定抗fgf7结构域抗体的表达、纯化和鉴定

背景:成纤维细胞生长因子(FGFs)参与血管生成、伤口愈合和胚胎发育。然而，成纤维细胞依赖性癌症中癌细胞生长的原因之一是成纤维细胞分泌的FGF7。因此，针对FGF7的抗体可用于治疗这些类型的癌症。方法:在以往的研究中，利用噬菌体展示技术已经鉴定出了一种具有单域抗体D53的噬菌体，该噬菌体具有抗人FGF7的功能。本研究制备并纯化了D53的分离形式。ELISA实验评价D53与FGF7的结合。通过对接研究和分子动力学(MD)模拟，探讨了D53-FGF7的相互作用模式。结果:western blotting和SDS-PAGE分析验证了表达和纯化过程。ELISA实验表明，D53能够识别并结合FGF7。对接研究和MD模拟表明，与虚拟VH相比，D53对FGF7具有更强的亲和力。结论:本研究的发现可用于FGF7抑制剂的生成和开发，并可能用于成纤维细胞依赖性癌症。

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分钟内获得全文求助全文

来源期刊