[Recombinant design and expression of human anti-rabies virus ScdsFv].
To constructed the recombinant human anti-rabies virus ScdsFv, cys sites were introduced into framework region (FR) of VH and VL genes which were amplified from human anti-rabies virus ScFv respectively using genetic point mutation technology. Cloned the ScdsFv gene into expression vector pET22b (+) and transformed into E. coli BL21 (DE3). The target protein was expressed by inducing with IPTG. Followed by renaturation in vitro and purified by Ni-NTA. The binding activity of ScdsFv was identified by Fluorescent antibody test (FAT) and ELISA. Results showed that recombinant ScdsFv were expressed at high level. Purity of the protein > 90% after purified by Ni-NTA and renaturaton in vitro. FAT and ELISA results demonstrated that ScdsFv could binding antigen specificity and was more stable than ScFv. Recombinant ScdsFv provided experiment materials for further functional study.
Cai
Kun
K
State Key Laboratory of Pathogens and Biosecurit, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China. ckreal@163.com
Wang
Hui
H
Bao
Shi-zhong
SZ
Shi
Jing
J
Hou
Xiao-jun
XJ
D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D015202:Protein Engineering; D011818:Rabies / Q000821:virology; D011820:Rabies virus / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins