Development of human antibody fragments using antibody phage display for the detection and diagnosis of Venezuelan equine encephalitis virus (VEEV).
Abstract: Venezuelan equine encephalitis virus (VEEV) belongs to the Alphavirus group. Several species of this family are also pathogenic to humans and are recognized as potential agents of biological warfare and terrorism. The objective of this work was the generation of recombinant antibodies for the detection of VEEV after a potential bioterrorism assault or an natural outbreak of VEEV. Results: In this work, human anti-VEEV single chain Fragments variable (scFv) were isolated for the first time from a human naïve antibody gene library using optimized selection processes. In total eleven different scFvs were identified and their immunological specificity was assessed. The specific detection of the VEEV strains TC83, H12/93 and 230 by the selected antibody fragments was proved. Active as well as formalin inactivated virus particles were recognized by the selected antibody fragments which could be also used for Western blot analysis of VEEV proteins and immunohistochemistry of VEEV infected cells. The anti-VEEV scFv phage clones did not show any cross-reactivity with Alphavirus species of the Western equine encephalitis virus (WEEV) and Eastern equine encephalitis virus (EEEV) antigenic complex, nor did they react with Chikungunya virus (CHIKV), if they were used as detection reagent. Conclusions: For the first time, this study describes the selection of antibodies against a human pathogenic virus from a human naïve scFv antibody gene library using complete, active virus particles as antigen. The broad and sensitive applicability of scFv-presenting phage for the immunological detection and diagnosis of Alphavirus species was demonstrated. The selected antibody fragments will improve the fast identification of VEEV in case of a biological warfare or terroristic attack or a natural outbreak.
Publication Date: 2008-09-02 PubMed ID: 18764933PubMed Central: PMC2543005DOI: 10.1186/1472-6750-8-66Google Scholar: Lookup
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Summary
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This research work focuses on developing human antibody fragments to detect and diagnose Venezuelan equine encephalitis virus (VEEV), which poses a significant threat in bio-warfare and terrorism. These fragments, known as single chain Fragments variable (scFv) were generated from a human naïve antibody gene library and were specifically used to detect both active and inactive particles of various VEEV strains, demonstrating their potential in prompt identification of VEEV in bio-warfare, terrorism, or natural outbreaks.
Development of Human Antibody Fragments
- The research was geared towards developing recombinant antibodies to detect the Venezuelan equine encephalitis virus (VEEV) which is classified under the Alphavirus species.
- The researchers generated single chain Fragments variable (scFv), a type of human antibody fragment, from a human naïve antibody gene library using optimized selection processes.
- Overall, eleven different scFv were created and evaluated for their immunological specificity.
Results: Specific Detection of VEEV Strains
- The developed antibody fragments were proven to specifically detect VEEV strains TC83, H12/93, and 230.
- The scFv could identify both active and formalin inactivated virus particles. This characteristic makes them valuable in various diagnostic procedures, including Western blot analysis of VEEV proteins and immunohistochemistry of VEEV-infected cells.
Testing Cross-Reactivity of the Anti-VEEV ScFv
- The researchers tested the anti-VEEV scFv phage clones for potential cross-reactivity with other Alphavirus species and Chikungunya virus (CHIKV). The results were negative, indicating that there was no cross-reactivity observed.
Conclusions and Potential Applications
- The study succeeded in producing, for the first time, antibodies against a human pathogenic virus from a human naïve scFv antibody gene library using complete, active virus particles as an antigen.
- The study proved the broad and sensitive utility of scFv-presenting phage in immunological detection and diagnosis of Alphavirus species.
- The developed scFv antibody fragments have potential to vastly improve the speed of VEEV identification in scenarios such as biological warfare, terroristic attacks, or natural outbreaks.
Cite This Article
APA
Kirsch MI, Hülseweh B, Nacke C, Rülker T, Schirrmann T, Marschall HJ, Hust M, Dübel S.
(2008).
Development of human antibody fragments using antibody phage display for the detection and diagnosis of Venezuelan equine encephalitis virus (VEEV).
BMC Biotechnol, 8, 66.
https://doi.org/10.1186/1472-6750-8-66 Publication
Researcher Affiliations
- Abteilung Biotechnologie, Institut für Biochemie und Biotechnologie, Technische Universität Braunschweig, Spielmannstrabetae 7, 38106, Braunschweig, Germany. martina.kirsch@tu-bs.de
MeSH Terms
- Biological Assay / methods
- Biosensing Techniques / methods
- Encephalitis Virus, Venezuelan Equine / immunology
- Encephalitis Virus, Venezuelan Equine / isolation & purification
- Humans
- Immunoassay / methods
- Immunoglobulin Fragments / analysis
- Immunoglobulin Fragments / immunology
- Peptide Library
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