Targeting of Adenovirus Serotype 5 Pseudotyped with Short Fiber from Serotype 41 to c-erbB2-Positive Cells using Bispecific Single-Chain Diabody

Elena A. Kashentseva, Joanne T. Douglas, Kurt R. Zinn, David T. Curiel, Igor P. Dmitriev

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Abstract: The purpose of the current study was to alter the broad native tropism of human adenovirus for virus targeting to c-erbB2-positive cancer cells. First, we engineered a single-chain antibody (scFv) against the c-erbB2 oncoprotein into minor capsid protein IX (pIX) of adenovirus serotype 5 (Ad5) in a manner commensurate with virion integrity and binding to the soluble extracellular c-erbB2 domain. To ablate native viral tropism and facilitate binding of the pIX-incorporated scFv to cellular c-erbB2, we replaced the Ad5 fiber with the Ad41 short (41s) fiber devoid of all known cell-binding determinants. The resultant Ad5F41sIX6.5 vector demonstrated increased cell binding and gene transfer as compared to the Ad5F41s control; however, this augmentation of virus infectivity was not c-erbB2 specific. Incorporation of a six-histidine (His6) peptide into the C-terminus of the 41s fiber protein resulted in markedly increased Ad5F41s6H infectivity in 293AR cells, which express a membrane-anchored scFv against the C-terminal oligohistidine tag, as compared to the Ad5F41s vector and the parental 293 cells. These data suggested that a 41s-fiber-incorporated His6 tag could serve for attachment of an adapter protein designed to guide Ad5F41s6H infection in a c-erbB2-specific manner. We therefore engineered a bispecific scFv diabody (scDb) combining affinities for both c-erbB2 and the His6 tag and showed its ability to provide up to 25-fold increase of Ad5F41s6H infectivity in c-erbB2-positive cells. Thus, Ad5 fiber replacement by a His6-tagged 41s fiber coupled with virus targeting mediated by an scDb adapter represents a promising strategy to confer Ad5 vector tropism for c-erbB2-positive cancer cells.

Original languageEnglish
Pages (from-to)443-461
Number of pages19
JournalJournal of Molecular Biology
Volume388
Issue number3
DOIs
StatePublished - May 8 2009

Keywords

  • adenovirus
  • c-erbB2
  • capsid protein IX
  • serotype 41 short fiber
  • single-chain diabody

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