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<p>Pulmonary fibrosis is a progressive disorder marked by excessive extracellular matrix deposition and decreased pulmonary function. Current available therapies like Nintedanib, provide limited efficacy and moderate therapeutic effect with notable side effects. Camelid nanobodies are famous due to their small size, high stability and target specificity, represent a novel therapeutic approach. In this study, a VHH nanobody library, generated against fibrosis was constructed from peripheral blood lymphocytes of immunized camels. The ~ 400&#xa0;bp VHH genes were cloned, expressed using phage display and purified via Ni-NTA chromatography. Therapeutic efficacy of the anti-fibrotic nanobody was evaluated in a bleomycin-induced pulmonary fibrosis in vivo mouse model (0.075 U/mL). This therapy resulted significant improvement in lung architecture and function, outperforming Nintedanib (60&#xa0;mg/kg). Along with lowered TNF-α levels, RT-PCR and immunoblotting revealed a 3.9 -fold and 4.1 -fold reduction in TGF-β and fibronectin expression respectively. This discovery and novel findings highlight camelid nanobodies as potent, non-immunogenic therapeutic candidates for pulmonary fibrosis, with promising applications in precision diagnostics and personalized medicine.</p> Graphical Abstract <p></p>

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Development and Optimization of Camelid Nanobody Libraries Using Molecular Cloning and Phage Display: Advancing Anti-fibrotic Therapy for Lung Fibrosis

  • 期刊论文

  • 发表日期：

    2026年6月16日

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  • Payal Pal,
  • Pramathadhip Paul,
  • Tania Debnath,
  • Avishek Das,
  • Annwaina Deb,
  • Paramita Barman,
  • Ena Ray Banerjee

摘要

Pulmonary fibrosis is a progressive disorder marked by excessive extracellular matrix deposition and decreased pulmonary function. Current available therapies like Nintedanib, provide limited efficacy and moderate therapeutic effect with notable side effects. Camelid nanobodies are famous due to their small size, high stability and target specificity, represent a novel therapeutic approach. In this study, a VHH nanobody library, generated against fibrosis was constructed from peripheral blood lymphocytes of immunized camels. The ~ 400 bp VHH genes were cloned, expressed using phage display and purified via Ni-NTA chromatography. Therapeutic efficacy of the anti-fibrotic nanobody was evaluated in a bleomycin-induced pulmonary fibrosis in vivo mouse model (0.075 U/mL). This therapy resulted significant improvement in lung architecture and function, outperforming Nintedanib (60 mg/kg). Along with lowered TNF-α levels, RT-PCR and immunoblotting revealed a 3.9 -fold and 4.1 -fold reduction in TGF-β and fibronectin expression respectively. This discovery and novel findings highlight camelid nanobodies as potent, non-immunogenic therapeutic candidates for pulmonary fibrosis, with promising applications in precision diagnostics and personalized medicine.

Graphical Abstract