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Tytuł pozycji:

Specific Binding of the Pathogenic Prion Isoform: Development and Characterization of a Humanized Single-Chain Variable Antibody Fragment.

Tytuł:
Specific Binding of the Pathogenic Prion Isoform: Development and Characterization of a Humanized Single-Chain Variable Antibody Fragment.
Autorzy:
Škrlj, Nives
Vranac, Tanja
Popović, Mara
Šerbec, Vladka Čurin
Dolinar, Marko
Temat:
PRION diseases
IMMUNOGLOBULINS
MONOCLONAL antibodies
IMMUNOHISTOCHEMISTRY
WESTERN immunoblotting
AFFINITY chromatography
SLOW virus diseases
ENZYME-linked immunosorbent assay
IMMUNE response
Źródło:
PLoS ONE; 2011, Vol. 6 Issue 1, p1-9, 9p
Czasopismo naukowe
Murine monoclonal antibody V5B2 which specifically recognizes the pathogenic form of the prion protein represents a potentially valuable tool in diagnostics or therapy of prion diseases. As murine antibodies elicit immune response in human, only modified forms can be used for therapeutic applications. We humanized a single-chain V5B2 antibody using variable domain resurfacing approach guided by computer modelling. Design based on sequence alignments and computer modelling resulted in a humanized version bearing 13 mutations compared to initial murine scFv. The humanized scFv was expressed in a dedicated bacterial system and purified by metal-affinity chromatography. Unaltered binding affinity to the original antigen was demonstrated by ELISA and maintained binding specificity was proved by Western blotting and immunohistochemistry. Since monoclonal antibodies against prion protein can antagonize prion propagation, humanized scFv specific for the pathogenic form of the prion protein might become a potential therapeutic reagent. [ABSTRACT FROM AUTHOR]
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