Abstract |
Hantaan virus (HTNV) is the type of Hantavirus causing hemorrhagic fever with renal syndrome, for which no specific therapeutics are available so far. Cell type-specific internalizing antibodies can be used to deliver therapeutics intracellularly to target cell and thus, have potential application in anti-HTNV infection. To achieve intracellular delivery of therapeutics, it is necessary to obtain antibodies that demonstrate sufficient cell type-specific binding, internalizing, and desired cellular trafficking. Here, we describe the prokaryotic expression, affinity purification, and functional testing of a single-chain Fv antibody fragment (scFv) against HTNV envelop glycoprotein (GP), an HTNV-specific antigen normally located on the membranes of HTNV-infected cells. This HTNV GP-targeting antibody, scFv3G1, was produced in the cytoplasm of Escherichia coli cells as a soluble protein and was purified by immobilized metal affinity chromatography. The purified scFv possessed a high specific antigen-binding activity to HTNV GP and HTNV-infected Vero E6 cells and could be internalized into HTNV-infected cells probably through the clathrin-dependent endocytosis pathways similar to that observed with transferrin. Our results showed that the E. coli-produced scFv had potential applications in targeted and intracellular delivery of therapeutics against HTNV infections.
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Authors | Jie Yang, Rui Chen, Junxia Wei, Fanglin Zhang, Yong Zhang, Lintao Jia, Yan Yan, Wen Luo, Yunxin Cao, Libo Yao, Jifeng Sun, Zhikai Xu, Angang Yang |
Journal | Applied microbiology and biotechnology
(Appl Microbiol Biotechnol)
Vol. 86
Issue 4
Pg. 1067-75
(Apr 2010)
ISSN: 1432-0614 [Electronic] Germany |
PMID | 20012277
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Recombinant Proteins
- Single-Chain Antibodies
- Viral Envelope Proteins
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Topics |
- Amino Acid Sequence
- Animals
- Base Sequence
- Chlorocebus aethiops
- Chromatography, Affinity
- Endocytosis
- Escherichia coli
(genetics)
- Hantaan virus
(immunology)
- Molecular Sequence Data
- Protein Binding
- Recombinant Proteins
(genetics, immunology, metabolism)
- Sequence Analysis, DNA
- Single-Chain Antibodies
(genetics, immunology, metabolism)
- Vero Cells
- Viral Envelope Proteins
(immunology)
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