Universität zu Köln

Friedel, Thorsten, Hanisch, Lydia J., Muth, Anke, Honegger, Annemarie ORCID: 0000-0002-3378-3967, Abken, Hinrich, Plueckthun, Andreas, Buchholz, Christian J. and Schneider, Irene C. (2015). Receptor-targeted lentiviral vectors are exceptionally sensitive toward the biophysical properties of the displayed single-chain Fv. Protein Eng. Des. Sel., 28 (4). S. 93 - 106. OXFORD: OXFORD UNIV PRESS. ISSN 1741-0134

Full text not available from this repository.

Abstract

An increasing number of applications require the expression of single-chain variable fragments (scFv) fusion proteins in mammalian cells at the cell surface membrane. Here we assessed the CD30-specific scFv HRS3, which is used in immunotherapy, for its ability to retarget lentiviral vectors (LVs) to CD30 and to mediate selective gene transfer into CD30-positive cells. Fused to the C-terminus of the type-II transmembrane protein hemagglutinin (H) of measles virus and expressed in LV packaging cells, gene transfer mediated by the released LV particles was inefficient. A series of point mutations in the scFv framework regions addressing its biophysical properties, which substantially improved production and increased the melting temperature without impairing its kinetic binding behavior to CD30, also improved the performance of LV particles. Gene transfer into CD30-positive cells increased similar to 100-fold due to improved transport of the H-scFv protein to the plasma membrane. Concomitantly, LV particle aggregation and syncytia formation in packaging cells were substantially reduced. The data suggest that syncytia formation can be triggered by trans-cellular dimerization of H-scFv proteins displayed on adjacent cells. Taken together, we show that the biophysical properties of the targeting ligand have a decisive role for the gene transfer efficiency of receptor-targeted LVs.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Friedel, Thorsten UNSPECIFIED UNSPECIFIED UNSPECIFIED Hanisch, Lydia J. UNSPECIFIED UNSPECIFIED UNSPECIFIED Muth, Anke UNSPECIFIED UNSPECIFIED UNSPECIFIED Honegger, Annemarie UNSPECIFIED orcid.org/0000-0002-3378-3967 UNSPECIFIED Abken, Hinrich UNSPECIFIED UNSPECIFIED UNSPECIFIED Plueckthun, Andreas UNSPECIFIED UNSPECIFIED UNSPECIFIED Buchholz, Christian J. UNSPECIFIED UNSPECIFIED UNSPECIFIED Schneider, Irene C. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-409363
DOI:	10.1093/protein/gzv005
Journal or Publication Title:	Protein Eng. Des. Sel.
Volume:	28
Number:	4
Page Range:	S. 93 - 106
Date:	2015
Publisher:	OXFORD UNIV PRESS
Place of Publication:	OXFORD
ISSN:	1741-0134
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language MEASLES-VIRUS HEMAGGLUTININ; IMMUNOGLOBULIN VARIABLE DOMAINS; V-H; ANTIBODY FRAGMENTS; ENDOTHELIAL-CELLS; GENE DELIVERY; PROTEIN; STABILITY; FRAMEWORK; ANTIGEN Multiple languages Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/40936