Generation and characterization of recombinant vascular targeting agents from hybridoma cell lines.

C. Gottstein, W. Wels, B. Ober, P. E. Thorpe

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

Vascular targeting agents (VTAs) can be produced by linking antibodies or antibody fragments directed against endothelial cell markers to effector moieties. So far, it has been necessary to produce the components of VTAs (antibody, antibody fragment, linker, and effector) separately and, subsequently, to conjugate them by biochemical reactions. We devised a cloning and expression system to allow rapid generation of recombinant VTAs from hybridoma cell lines. The VTAs consist of a single chain Fv antibody fragment as a targeting moiety and either truncated Pseudomonas exotoxin (resulting in immunotoxins) or truncated human tissue factor (resulting in coaguligands) as effectors. The system was applied to generate recombinant immunotoxins and coaguligands directed against endoglin, vascular endothelial growth factor (VEGF):VEGF receptor (VEGFR) complex and vascular cell adhesion molecule 1 (VCAM-1). The fusion proteins exhibited similar functional activity to analogous biochemical constructs. This is the first report to describe the generation and characterization of recombinant coaguligands.

Original languageEnglish (US)
Pages (from-to)190-194, 196, 198 passim
JournalBioTechniques
Volume30
Issue number1
StatePublished - Jan 2001

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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    Gottstein, C., Wels, W., Ober, B., & Thorpe, P. E. (2001). Generation and characterization of recombinant vascular targeting agents from hybridoma cell lines. BioTechniques, 30(1), 190-194, 196, 198 passim.