[Improved biochemical characteristics of human disulfide-stabilized Fv fragment to rabies virus].
This study is (1) to improve the stabilization of human scFv to rabies virus; (2) to prepare active human dsFv fragment; and (3) to evaluate the biological activities of dsFv. The dsFv V(H) and VL were separately expressed in PET22b(+)/BL21 (DE3), solublized and combined in appropriate molar ratio in refolding solution. The resultant dsFv fragments were evaluated for its protection against rabies virus, its affinity and stability, in reference to the cognate scFv. The dsFv was found to bind specifically to Vero vaccine of rabies virus. Compared to the scFv, the dsFv was more stable, had higher affinity, and was able to inhibit the infection of Rabies virus to Vero cell. This established a solid basis for the clinical application of dsFv to rabies virus.
Zhao
Xiao-Ling
XL
Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China.
Yin
Jun
J
Zhang
Su-Juan
SJ
Chen
Wei-Qiang
WQ
D000914:Antibodies, Viral / Q000276:immunology*; D004220:Disulfides / Q000737:chemistry; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology* / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000276:immunology* / Q000378:metabolism; D011820:Rabies virus / Q000276:immunology*
D000914:Antibodies, Viral; D004220:Disulfides; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; C055966:immunoglobulin Fv