Lung-Targeting Nanobodies Against Human Pulmonary Surfactant Protein A and a Method for Producing the Same

摘要

The present invention relates to the field of biochemistry and pharmaceutical technologies. The present invention provides nanobodies that bind to human pulmonary surfactant protein A (SP-A) as well as the preparing methods and use of the same. The nanobody comprise an amino acid sequence having the formula of Q(x)2LVESGG(x)2V (x)2G(x) SL(x) LS(x)24E (x)n2KG(x)4S(x)n3T(x)2Y(x) C(x)n4S(x)n5V(x)n6R; wherein x is any amino acid; n2˜n6 are positive integers; 1≦n2≦21; 1≦n3≦19; 1≦n4≦50; 1≦n5≦22; 1≦n6≦8. The present invention take fresh frozen sections of lung as antigen, gene sequences with high affinity with hSP-A were obtained by constructing an SP-A antibody library and affinity selection, and nanobodies with high affinity and small molecule weight were obtained by induced expression of the gene sequences through a prokaryotic expression vector. Immunohistochemistry and in vivo imaging in nude mice showed the nanobodies have high specificity for targeting lung tissue.

主权项

1. A human lung-targeting nanobody, comprising an amino acid sequence having the formula of Q(x)2LVESGG(x)2V (x)2G(x) SL(x) LS(x)24E (x)n2KG(x)4S(x)n3T(x)2Y(x) C(x)n4S(x)n5V(x)n6R; wherein x is any amino acid; n2˜n6 are positive integers; 1≦n2≦21; 1≦n3≦19; 1≦n4≦50; 1≦n5≦22; 1≦n6≦8.