Nanocell targeting using engineered bispecific antibodies.

Taylor, Karin, Howard, Christopher B., Jones, Martina L., Sedliarou, Ilya, MacDiarmid, Jennifer, Brahmbhatt, Himanshu, Munro, Trent P. and Mahler, Stephen M. (2014) Nanocell targeting using engineered bispecific antibodies.. mAbs, 7 1: 53-65. doi:10.4161/19420862.2014.985952

Author Taylor, Karin
Howard, Christopher B.
Jones, Martina L.
Sedliarou, Ilya
MacDiarmid, Jennifer
Brahmbhatt, Himanshu
Munro, Trent P.
Mahler, Stephen M.
Title Nanocell targeting using engineered bispecific antibodies.
Journal name mAbs   Check publisher's open access policy
ISSN 1942-0862
Publication date 2014-12
Year available 2014
Sub-type Article (original research)
DOI 10.4161/19420862.2014.985952
Open Access Status
Volume 7
Issue 1
Start page 53
End page 65
Total pages 13
Place of publication London, United Kingdom
Publisher Taylor and Fancis
Language eng
Formatted abstract
There are many design formats for bispecific antibodies (BsAbs), and the best design choice is highly dependent on the final application. Our aim was to engineer BsAbs to target a novel nanocell (EnGeneIC Delivery Vehicle or EDVTMnanocell) to the epidermal growth factor receptor (EGFR). EDVTMnanocells are coated with lipopolysaccharide (LPS), and BsAb designs incorporated single chain Fv (scFv) fragments derived from an anti-LPS antibody (1H10) and an anti-EGFR antibody, ABX-EGF. We engineered various BsAb formats with monovalent or bivalent binding arms and linked scFv fragments via either glycine-serine (G4S) or Fc-linkers. Binding analyses utilizing ELISA, surface plasmon resonance, bio-layer interferometry, flow cytometry and fluorescence microscopy showed that binding to LPS and to either soluble recombinant EGFR or MDA-MB-468 cells expressing EGFR, was conserved for all construct designs. However, the Fc-linked BsAbs led to nanocell clumping upon binding to EDVTMnanocells. Clumping was eliminated when additional disulfide bonds were incorporated into the scFv components of the BsAbs, but this resulted in lower BsAb expression. The G4S-linked tandem scFv BsAb format was the optimal design with respect to EDV binding and expression yield. Doxorubicin-loaded EDVTMnanocells actively targeted with tandem scFv BsAb in vivo to MDA-MB-468-derived tumors in mouse xenograft models enhanced tumor regression by 40% compared to passively targeted EDVTMnanocells. BsAbs therefore provide a functional means to deliver EDVTMnanocells to target cells.
Keyword Bispecific antibody
Bispecific antibody
Disulfide Bridges
Mammalian expression
Monoclonal antibody
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

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Created: Thu, 15 Jan 2015, 09:14:13 EST by Cathy Fouhy on behalf of Aust Institute for Bioengineering & Nanotechnology