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[Recombinant design and expression of human anti-rabies virus ScdsFv].
- Source :
-
Wei sheng wu xue bao = Acta microbiologica Sinica [Wei Sheng Wu Xue Bao] 2007 Aug; Vol. 47 (4), pp. 673-6. - Publication Year :
- 2007
-
Abstract
- 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.
- Subjects :
- Antibodies, Monoclonal immunology
Antibodies, Monoclonal isolation & purification
Cloning, Molecular
Escherichia coli genetics
Escherichia coli metabolism
Genetic Vectors genetics
Humans
Immunoglobulin Variable Region immunology
Immunoglobulin Variable Region isolation & purification
Rabies virology
Rabies virus genetics
Recombinant Proteins genetics
Recombinant Proteins immunology
Recombinant Proteins isolation & purification
Antibodies, Monoclonal genetics
Gene Expression
Immunoglobulin Variable Region genetics
Protein Engineering
Rabies virus immunology
Subjects
Details
- Language :
- Chinese
- ISSN :
- 0001-6209
- Volume :
- 47
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Wei sheng wu xue bao = Acta microbiologica Sinica
- Publication Type :
- Academic Journal
- Accession number :
- 17944370