Monoclonal antibody–polyethyleneimine conjugates targeting Her-2/ neu or CD90 allow cell type-specific nonviral gene delivery

• Published in: Journal of controlled release 2005-02, Vol.102 (3), p.737-747
• Main Authors: Strehblow, Claudia, Schuster, Maj, Moritz, Thomas, Kirch, Hans-Christoph, Opalka, Bertram, Petri, Jean Bernhard
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - administration & dosage; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - genetics; Antibodies, Monoclonal, Humanized; Biological and medical sciences; Cells, Cultured; Gene Targeting - methods; Gene transfer; General pharmacology; Genes, erbB-2 - genetics; Her-2/ neu (Trastuzumab); Humans; Medical sciences; Mice; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Polyethyleneimine - administration & dosage; Polyethyleneimine - chemistry; Positional targeting; Protein Binding - drug effects; Thy-1; Thy-1 Antigens - genetics; Transfection - methods; Trastuzumab
• ISSN: 0168-3659; 1873-4995
• DOI: 10.1016/j.jconrel.2004.10.033

We have developed an efficient and cell-specific nonviral gene delivery system using monoclonal antibodies (mAb) coupled with branched 25-kDa polyethyleneimine (PEI). The system was evaluated for two model antibodies with different well-characterized antigen specificities: the mouse anti-human IgG1 mAb AS02 recognizing human CD90 (hThy-1) which is expressed on human fibroblasts, and the humanized anti-Her-2/ neu mAb Trastuzumab recently introduced for the treatment of Her-2/ neu-positive breast cancer. Efficacy and selectivity of gene delivery were evaluated for covalent mAb–PEI conjugates coupled with N-succinimidyl-3-(2-pyridyldithio)proprionate (SPDP) or N-succinimidyl-4-(maleimidomethyl)-cyclohexancarboxylate (SMCC), or, as a newly introduced coupling reagent, noncovalent complexes of mAb with 3-(2-(2-(vinylsulfonyl)ethylthio)ethyl)quinazoline-2,4(1 H,3 H)-dione (IBFB 110001) coupled to PEI. An enhanced green fluorescent protein (EGFP)-expressing reporter plasmid was used to monitor transfection efficiencies of cell lines expressing different levels of hCD90 or Her-2/ neu. While mAb–PEI conjugates coupled with SPDP resulted in antigen-nonspecific EGFP expression, conjugates coupled with SMCC or IBFB 110001 enabled antigen-specific gene delivery. Thus, Her-2/ neu–PEI conjugates prepared with IBFB 110001 allowed to transfect 23±2% of Her-2/ neu-positive SKOV-3 versus 0.5±2% of Her-2/ neu-negative MB-468 cells. Proof of principle for specific antibody-mediated gene transfer was demonstrated by saturating the Her-2/ neu receptor with free anti-Her-2/ neu, thereby blocking subsequent transfection with anti-Her-2/ neu–PEI/DNA complexes.