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Nanocell targeting using engineered bispecific antibodies.

Karin Taylor1, Christopher B Howard, Martina L Jones

  • 1a Australian Institute for Bioengineering and Nanotechnology (AIBN) ; University of Queensland, St Lucia ; Queensland , Australia.

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|December 20, 2014
PubMed
Summary
This summary is machine-generated.

Engineered bispecific antibodies (BsAbs) effectively target EnGeneIC Delivery Vehicles (EDVs) to epidermal growth factor receptor (EGFR) expressing cells. The optimal G4S-linked tandem scFv BsAb format enhanced tumor regression in vivo.

Keywords:
BsAb, bispecific antibodyEDVTM, EnGeneIC Delivery VehicleEGFR, epidermal growth factor receptorIgG, immunoglobulin GLPS, lipopolysaccharideNP, nanoparticlebispecific antibodydisulfide bridgemAb, monoclonal antibodymammalian expressionnanoparticlescFv, single chain variable fragmentsingle chain Fvsurface plasmon resonancetumor regression

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Area of Science:

  • Biotechnology
  • Antibody Engineering
  • Nanomedicine

Background:

  • Bispecific antibodies (BsAbs) offer versatile therapeutic strategies.
  • EnGeneIC Delivery Vehicles (EDVs) are novel nanocell constructs.
  • Targeting EDVs to specific receptors like EGFR is crucial for precision medicine.

Purpose of the Study:

  • To engineer and evaluate various bispecific antibody (BsAb) formats for targeting EDVs to the epidermal growth factor receptor (EGFR).
  • To identify an optimal BsAb design balancing EDV binding, expression yield, and in vivo efficacy.
  • To assess the therapeutic potential of targeted EDVs in a preclinical cancer model.

Main Methods:

  • Engineered BsAbs with different formats (monovalent/bivalent arms, G4S/Fc linkers) using anti-LPS and anti-EGFR scFv fragments.
  • Evaluated BsAb binding to EDVs and EGFR-expressing cells using various assays (ELISA, SPR, BLI, flow cytometry, microscopy).
  • Assessed in vivo tumor regression of doxorubicin-loaded EDVs targeted with the optimal BsAb in mouse xenograft models.

Main Results:

  • All engineered BsAb formats demonstrated binding to EDVs and EGFR.
  • Fc-linked BsAbs caused undesirable nanocell clumping; disulfide bond incorporation reduced clumping but lowered expression.
  • The G4S-linked tandem scFv BsAb format showed optimal EDV binding and expression yield.
  • Active targeting with BsAb-conjugated EDVs resulted in a 40% enhancement in tumor regression compared to passive targeting.

Conclusions:

  • BsAb design significantly impacts EDV targeting and therapeutic efficacy.
  • G4S-linked tandem scFv BsAbs represent an optimal format for EGFR-targeted EDV delivery.
  • BsAbs provide a functional strategy for actively targeting nanocell drug delivery systems to tumors, enhancing therapeutic outcomes.