PMID: 8581376Oct 1, 1995Paper

Properties of bispecific single chain antibodies expressed in Escherichia coli

Journal of Hematotherapy
D M KranzE R Wilson

Abstract

Single chain bispecific antibodies, in which the genes that encode the VH and VL regions are linked in tandem, may offer some advantages over other methods of bispecific antibody preparation. To begin to evaluate the potential of this system, a single chain bispecific antibody (scFv2) that binds to the T cell receptor of a cytotoxic T cell clone and to the hapten fluorescein was constructed. The individual scFv regions were joined by a 25 amino acid linker, and the scFv2 protein was obtained from insoluble inclusion bodies after guanidine solubilization and refolding. Fluorescein-purified scFv2 is active at concentrations of approximately 10 nM (approximately 1 microgram/ml) and above in mediating lysis of fluorescein-coupled tumor cells by the cytotoxic T lymphocytes (CTL). This system has now been used to evaluate various features of the scFv2 approach: (a) affinities of the scFv2 for the TCR and fluorescein, (b) yields of the scFv2 from several different purification schemes, (c) relationship of antigen density (i.e., fluorescein density on tumor cells) to the ability to redirect lysis of the tumor cell, and (d) relationship of scFv2 affinity for the tumor antigen to the ability to redirect lysis. This study was performed by...Continue Reading

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Mar 6, 1992·Cell·P MombaertsV E Papaioannou
Oct 21, 1988·Science·R E BirdM Whitlow
Dec 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·D M KranzH N Eisen
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Jul 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·P HolligerG Winter

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Citations

Feb 5, 2003·Advanced Drug Delivery Reviews·Ying Cao, Laura Lam
Jun 1, 1997·Immunotechnology : an International Journal of Immunological Engineering·A Plückthun, P Pack
Aug 1, 2000·Immunology Today·M LittleG Moldenhauer
May 29, 2007·Oncogene·I Zafir-LavieY Reiter
May 9, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·A M McCallL M Weiner
Nov 17, 1998·Bioconjugate Chemistry·Y Cao, M R Suresh

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