Scholarlink

Background <p>Although nanobodies have superior characteristics to conventional antibodies, their affinities are often lower because of a single interaction site for antigen binding. Therefore, in the current study, a bioinformatics approach was employed to enhance the affinity of a humanized anti-VEGF nanobody even that of the commercially available antibody, bevacizumab.</p> Methods <p>The in vitro safe dose and the affinity of the wild and mutated nanobodies were determined and compared to bevacizumab. The best mutated nanobodies (MHNb136 and MHNb256) were selected by implementing umbrella sampling simulation to improve the binding affinity of the wild-type nanobody (WHNb) to VEGF. In vitro safety investigations were performed on nanobodies with doses of 12.5, 25, 50, and 100 µg, equivalent to the dose of bevacizumab as the conventional treatment.</p> Results <p>Ultimately, umbrella sampling results showed that MHNb136 and MHNb256 bind to VEGF more strongly, with <i>ΔG</i> values of −39.3 and −32.64 kcal/mol, respectively, than WHNb, which has a <i>ΔG</i> value of −19.05 kcal/mol. BIACORE assay results indicated that MHNb136 and MHNb256 bind to VEGF with higher affinity than bevacizumab as a reference.</p> Conclusion <p>Eventually, MHNb136 and MHNb256 can be subjected for further clinical investigation due to the superiority to bevacizumab.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Affinity enhancement of an anti-VEGF humanized nanobody as a superior candidate to bevacizumab: integrative in silico and in vitro studies

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年5月12日

查看全文
  • Mir Salar Kazemi,
  • Seyed Shahriar Arab,
  • Fatemeh Kazemi-Lomedasht,
  • Reza Ahangari Cohan,
  • Fatemeh Zandi,
  • Alireza Shoari,
  • Mohammad Hosseininejad-Chafi,
  • Golnoosh Mahjoobi,
  • Arghavan Ashja Ardalan,
  • Mozhgan Rezaei Kanavi,
  • Mahdi Behdani

摘要

Background

Although nanobodies have superior characteristics to conventional antibodies, their affinities are often lower because of a single interaction site for antigen binding. Therefore, in the current study, a bioinformatics approach was employed to enhance the affinity of a humanized anti-VEGF nanobody even that of the commercially available antibody, bevacizumab.

Methods

The in vitro safe dose and the affinity of the wild and mutated nanobodies were determined and compared to bevacizumab. The best mutated nanobodies (MHNb136 and MHNb256) were selected by implementing umbrella sampling simulation to improve the binding affinity of the wild-type nanobody (WHNb) to VEGF. In vitro safety investigations were performed on nanobodies with doses of 12.5, 25, 50, and 100 µg, equivalent to the dose of bevacizumab as the conventional treatment.

Results

Ultimately, umbrella sampling results showed that MHNb136 and MHNb256 bind to VEGF more strongly, with ΔG values of −39.3 and −32.64 kcal/mol, respectively, than WHNb, which has a ΔG value of −19.05 kcal/mol. BIACORE assay results indicated that MHNb136 and MHNb256 bind to VEGF with higher affinity than bevacizumab as a reference.

Conclusion

Eventually, MHNb136 and MHNb256 can be subjected for further clinical investigation due to the superiority to bevacizumab.