The Study Of Lung-targeting Anti-hSPA Nanobodies

Objectives: This thesis with pulmonary surfactant protein A(SP- A) as the target, the preparation of nano antibodies against human lung SP- A, with clinical medicine provides the ideal targeting ligands.This thesis with pulmonary surfactant protein A(SP- A) as the target, the preparation of nano antibodies against human lung SP- A, with clinical medicine provides the ideal targeting ligands.Methods:1. The alveolar surfactant protein A(h SP- A) acquisition and identification:from fresh human lung tissue using animal protein extract extracted proteins of lung tissue,Westernblot andspecificity.2. Lung tissue SP- A nano antibody specificity(h SPA- Nb) screening: by using the solid-phase method of phage antibody library for three rounds of competive, get enrichment of tertiary screening antibody libraries, and the indirect phage- ELISA from highly specific gene sequences of triple antibody library.3. The expression and purification of specific h SPA- Nb: positive strains and sequencing analysis to construct recombinant plasmid PET- 26 b(+)/Nb4- whether,further purification.4. h SPA, Nb4 lung targeting and safety inspection: using Western Blot and El ISA to detect protein specificity, cell fluorescence experiment, immunohistochemical and mice in vivo imaging of lung targeting property and safety.Results: This study first success was prepared and the lung tissue h SP- there is A specific combination of h SP resistance- A nano antibody Nb4.Conclusions: The nano antibodies can not only h SP- A specific binding, also can be implemented by targeting natural h SP- A highly specific lung targeting.