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[Preparation and bioactivity of anti-human red blood cell ScFv and CSFV E2 bifunctional fusion protein].
- Source :
-
Sheng wu gong cheng xue bao = Chinese journal of biotechnology [Sheng Wu Gong Cheng Xue Bao] 2010 Jan; Vol. 26 (1), pp. 28-34. - Publication Year :
- 2010
-
Abstract
- The aim of this study is to construct a bifunctional fusion protein, which can conjugate both human red blood cells and antibodies against classical swine fever virus (CSFV). We respectively amplified 2E8ScFv and mE2 genes from different recombinant vectors, in which 2E8ScFv gene is the single chain Fv gene against H antigen of human red blood cells, whereas mE2 gene is the main antigen coding region gene of CSFV E2 protein. We used overlap extension PCR to obtain an artificial fusion gene segment 2E8mE2 containing genes of Both 2E8ScFv and mE2, then ligated into the expression vector pET-DsbA and expressed in Escherichia coli BL21(DE3) PlysS host cells, after induced with IPTG the target fusion protein was successfully expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. We purified the fusion protein and renatured it from inclusion bodies to obtain a native state of well biological activity. The Erythrocyte agglutination test results indicated that the fusion protein can conjugate both human red blood cells and antibodies of CSFV.
- Subjects :
- Escherichia coli genetics
Escherichia coli metabolism
Humans
Immunoglobulin Fragments genetics
Immunoglobulin Fragments immunology
Immunoglobulin Variable Region biosynthesis
Immunoglobulin Variable Region immunology
Recombinant Fusion Proteins biosynthesis
Recombinant Fusion Proteins genetics
Single-Chain Antibodies genetics
Viral Envelope Proteins genetics
Erythrocytes immunology
Immunoglobulin Fragments biosynthesis
Immunoglobulin Variable Region genetics
Single-Chain Antibodies biosynthesis
Viral Envelope Proteins biosynthesis
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1000-3061
- Volume :
- 26
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Sheng wu gong cheng xue bao = Chinese journal of biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 20353089