Binding characteristics and tumor targeting of a covalently linked divalent CC49 single-chain antibody

Guy W. Beresford, Gabriela Pavlinkova, Barbara J.M. Booth, Surinder K. Batra, David Colcher

Research output: Contribution to journalArticlepeer-review

43 Scopus citations

Abstract

Multivalency is a recognized means of increasing the functional affinity of single-chain Fvs (scFvs) for optimizing tumor uptake. A unique divalent single-chain Fv protein [sc(Fv)2], based on the variable regions of the monoclonal antibody (MAb) CC49, has been generated that differs from other dimeric single-chain constructs in that a linker sequence (L) is encoded between the repeated V(L) and V(H) domains (V(L)-L-V(H)L-V(L)-L-V(H)). This construct was expressed in soluble form in Escherichia call and purified by ion-exchange and gel-filtration chromatography. Purity and immunoreactivity were determined by SDS-PAGE, HPLC and competitive RIA. sc(Fv)2 exhibited a relative K(A) (3.34 x 107 M-1) similar to that of the native IgG (1.14 x 108 M-1) as determined by BIAcore analysis. Pharmacokinetic studies showed rapid blood clearance for sc(Fv)2, with a T(1/2) less than 40 min. Whole- body clearance analysis also revealed rapid clearance, suggesting no significant retention in the extravascular space or normal tissues. Biodistribution studies of radiolabeled sc(Fv)2 showed tumor uptake greater than 6% ID/g after 30 min, which remained at this level for 6 hr. High tumor uptake and retention of sc(Fv)2 coupled with rapid blood and whole-body clearance makes this dimeric scFv of MAb CC49 a strong candidate for imaging and therapeutic applications.

Original languageEnglish (US)
Pages (from-to)911-917
Number of pages7
JournalInternational Journal of Cancer
Volume81
Issue number6
DOIs
StatePublished - 1999

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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