Related Experiment Videos
[Recombinant antibodies to tick-borne encephalitis virus]
Voprosy Virusologii
|January 14, 2003
Summary
Researchers engineered dimeric single-chain variable fragments (scFv) for tick-borne encephalitis virus glycoprotein E. These scFv antibodies, produced in E. coli, demonstrate specific binding to the virus, advancing diagnostic tools.
Area of Science:
- Biotechnology
- Molecular Biology
- Immunology
Context:
- Tick-borne encephalitis (TBE) virus poses a significant public health threat.
- Development of effective diagnostic tools for TBE is crucial for disease management.
- Single-chain variable fragments (scFv) offer potential for targeted therapeutic and diagnostic applications.
Purpose:
- To design and produce dimeric single-chain variable fragments (scFv) targeting tick-borne encephalitis virus glycoprotein E.
- To characterize the specificity and binding capabilities of engineered scFv antibodies.
- To confirm the epitope recognized by the scFv antibodies and compare it with parental monoclonal antibodies.
Summary:
- An artificial gene for dimeric scFv targeting TBE virus glycoprotein E was designed based on a monomeric scFv gene.
- Histidine tag sequences were added to facilitate purification of both monomeric and dimeric scFv antibodies.
- Escherichia coli strains were developed for antibody production, followed by purification using Ni-NTA resin.
- ELISA and Western-blot analyses confirmed the specific binding of purified scFv antibodies to TBE virus.
- Competitive immunoassay validated that the scFv antibodies recognize the same epitope on glycoprotein E as the E6B monoclonal antibody.
Impact:
- Successful production of functional dimeric scFv antibodies against TBE virus glycoprotein E.
- Demonstrated specificity of engineered antibodies, paving the way for improved diagnostic assays.
- Provides a foundation for developing novel diagnostic or therapeutic strategies for tick-borne encephalitis.