Autor(es): Kirsch Martina Inga, Hülseweh Birgit, Nacke Christoph, Rülker Torsten, Schirrmann Thomas, Marschall Hans-Jürgen, Hust Michael, Dübel Stefan

Resumo: Venezuelan equine encephalitis virus (VEEV) belongs to the Alphavirus group. Several species of this family are also pathogenic to humans - are recognized as potential agents of biological warfare - 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. 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 - their immunological specificity was assessed. The specific detection of the VEEV strains TC83, H12/93 - 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 - 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) - Eastern equine encephalitis virus (EEEV) antigenic complex, nor did they react with Chikungunya virus (CHIKV), if they were used as detection reagent. 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 - sensitive applicability of scFv-presenting phage for the immunological detection - 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.

Imprenta: BMC Biotechnology, v. 8, p. 66, 2008

Identificador do objeto digital: 10.1186/1472-6750-8-66

Descritores: Chikungunya virus - Cell ; Chikungunya virus - Molecular structure ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Proteins ; Chikungunya virus - Antibodies ; Chikungunya Virus - Virus ; Chikungunya virus - Immunology

Data de publicação: 2008

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