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A novel nanobody specific for respiratory surfactant protein A has potential for lung targeting.

Wang SM, He X, Li N, Yu F, Hu Y, Wang LS, Zhang P, Du YK, Du SS, Yin ZF, Wei YR, Mulet X, Coia G, Weng D, He JH, Wu M, Li HP - Int J Nanomedicine (2015)

Bottom Line: Here, we generated recombinant rat pulmonary SPA (rSPA) as an antigen and immunized an alpaca to produce two nanobodies (the smallest naturally occurring antibodies) specific for rSPA, designated Nb6 and Nb17.Using immunohistochemistry, we demonstrated that these anti-rSPA nanobodies selectively bound to rat lungs with high affinity.Collectively, we have generated and characterized lung-specific nanobodies, which may be applicable for lung drug delivery.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, People's Republic of China.

ABSTRACT
Lung-targeting drugs are thought to be potential therapies of refractory lung diseases by maximizing local drug concentrations in the lung to avoid systemic circulation. However, a major limitation in developing lung-targeted drugs is the acquirement of lung-specific ligands. Pulmonary surfactant protein A (SPA) is predominantly synthesized by type II alveolar epithelial cells, and may serve as a potential lung-targeting ligand. Here, we generated recombinant rat pulmonary SPA (rSPA) as an antigen and immunized an alpaca to produce two nanobodies (the smallest naturally occurring antibodies) specific for rSPA, designated Nb6 and Nb17. To assess these nanobodies' potential for lung targeting, we evaluated their specificity to lung tissue and toxicity in mice. Using immunohistochemistry, we demonstrated that these anti-rSPA nanobodies selectively bound to rat lungs with high affinity. Furthermore, we intravenously injected fluorescein isothiocyanate-Nb17 in nude mice and observed its preferential accumulation in the lung to other tissues, suggesting high affinity of the nanobody for the lung. Studying acute and chronic toxicity of Nb17 revealed its safety in rats without causing apparent histological alterations. Collectively, we have generated and characterized lung-specific nanobodies, which may be applicable for lung drug delivery.

No MeSH data available.


Related in: MedlinePlus

Biophysical analyses of Nb17.Notes: Antigen-specific ELISA was performed to test the binding ability of Nb17, SPA-poly-ant at various temperatures (A) and at various pH (B).Abbreviations: ELISA, enzyme-linked immunosorbent assay; SPA-poly-ant, surfactant protein A polyclonal antibody.
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f5-ijn-10-2857: Biophysical analyses of Nb17.Notes: Antigen-specific ELISA was performed to test the binding ability of Nb17, SPA-poly-ant at various temperatures (A) and at various pH (B).Abbreviations: ELISA, enzyme-linked immunosorbent assay; SPA-poly-ant, surfactant protein A polyclonal antibody.

Mentions: Since Nb17 showed a higher binding ability to rSPA in immunohistochemistry than Nb6, we chose Nb17 to evaluate its targeting ability further. In order to investigate the biophysical feature of Nb17, a standard antigen-specific ELISA was performed to test whether binding could be observed at various temperatures and various pH values. As shown in Figure 5, contrary to conventional rSPA-poly-ant, Nb17 was found to be far more stable. In more detail, for Nb17, it is clearly shown that it specifically bound to rSPA even at temperatures of 90°C and pH values of 3.5 or at 8.0. However, rSPA-poly-ant showed almost no antigen binding when the temperature was above 70°C and acid or alkaline conditions were extreme.


A novel nanobody specific for respiratory surfactant protein A has potential for lung targeting.

Wang SM, He X, Li N, Yu F, Hu Y, Wang LS, Zhang P, Du YK, Du SS, Yin ZF, Wei YR, Mulet X, Coia G, Weng D, He JH, Wu M, Li HP - Int J Nanomedicine (2015)

Biophysical analyses of Nb17.Notes: Antigen-specific ELISA was performed to test the binding ability of Nb17, SPA-poly-ant at various temperatures (A) and at various pH (B).Abbreviations: ELISA, enzyme-linked immunosorbent assay; SPA-poly-ant, surfactant protein A polyclonal antibody.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4403696&req=5

f5-ijn-10-2857: Biophysical analyses of Nb17.Notes: Antigen-specific ELISA was performed to test the binding ability of Nb17, SPA-poly-ant at various temperatures (A) and at various pH (B).Abbreviations: ELISA, enzyme-linked immunosorbent assay; SPA-poly-ant, surfactant protein A polyclonal antibody.
Mentions: Since Nb17 showed a higher binding ability to rSPA in immunohistochemistry than Nb6, we chose Nb17 to evaluate its targeting ability further. In order to investigate the biophysical feature of Nb17, a standard antigen-specific ELISA was performed to test whether binding could be observed at various temperatures and various pH values. As shown in Figure 5, contrary to conventional rSPA-poly-ant, Nb17 was found to be far more stable. In more detail, for Nb17, it is clearly shown that it specifically bound to rSPA even at temperatures of 90°C and pH values of 3.5 or at 8.0. However, rSPA-poly-ant showed almost no antigen binding when the temperature was above 70°C and acid or alkaline conditions were extreme.

Bottom Line: Here, we generated recombinant rat pulmonary SPA (rSPA) as an antigen and immunized an alpaca to produce two nanobodies (the smallest naturally occurring antibodies) specific for rSPA, designated Nb6 and Nb17.Using immunohistochemistry, we demonstrated that these anti-rSPA nanobodies selectively bound to rat lungs with high affinity.Collectively, we have generated and characterized lung-specific nanobodies, which may be applicable for lung drug delivery.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, People's Republic of China.

ABSTRACT
Lung-targeting drugs are thought to be potential therapies of refractory lung diseases by maximizing local drug concentrations in the lung to avoid systemic circulation. However, a major limitation in developing lung-targeted drugs is the acquirement of lung-specific ligands. Pulmonary surfactant protein A (SPA) is predominantly synthesized by type II alveolar epithelial cells, and may serve as a potential lung-targeting ligand. Here, we generated recombinant rat pulmonary SPA (rSPA) as an antigen and immunized an alpaca to produce two nanobodies (the smallest naturally occurring antibodies) specific for rSPA, designated Nb6 and Nb17. To assess these nanobodies' potential for lung targeting, we evaluated their specificity to lung tissue and toxicity in mice. Using immunohistochemistry, we demonstrated that these anti-rSPA nanobodies selectively bound to rat lungs with high affinity. Furthermore, we intravenously injected fluorescein isothiocyanate-Nb17 in nude mice and observed its preferential accumulation in the lung to other tissues, suggesting high affinity of the nanobody for the lung. Studying acute and chronic toxicity of Nb17 revealed its safety in rats without causing apparent histological alterations. Collectively, we have generated and characterized lung-specific nanobodies, which may be applicable for lung drug delivery.

No MeSH data available.


Related in: MedlinePlus