Targeting a recombinant adenovirus vector to HCC cells using a bifunctional Fab-antibody conjugate.
We developed a specific adenoviral gene delivery system with monoclonal antibody (mAb) AF-20 that binds to a 180 kDa antigen highly expressed on human hepatocellular carcinoma (HCC) cells. A bifunctional Fab-antibody conjugate (2Hx-2-AF-20) was generated through AF-20 mAb crosslinkage to an anti-hexon antibody Fab fragment. Uptake of adenoviral particles and gene expression was examined in FOCUS HCC and NIH 3T3 cells by immunofluorescence; beta-galactosidase expression levels were determined following competitive inhibition of adenoviral CAR receptor by excess fibre knob protein. The chimeric complex was rapidly internalized at 37 degrees C, and enhanced levels of reporter gene expression was observed in AF-20 antigen positive HCC cells, but not in AF-20 antigen negative NIH 3T3 control cells. Targeting of recombinant adenoviral vectors to a tumor associated antigen by a bifunctional Fab-antibody conjugate is a promising approach to enhance specificity and efficiency of gene delivery to HCC.
Yoon
S K
SK
Molecular Hepatology Laboratory, Massachusetts General Hospital Cancer Center, Charlestown, MA 02129, USA.
Mohr
L
L
O'Riordan
C R
CR
Lachapelle
A
A
Armentano
D
D
Wands
J R
JR
D016475:3T3 Cells; D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D000912:Antibodies, Neoplasm / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D000914:Antibodies, Viral / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D001667:Binding, Competitive; D002213:Capsid / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D036022:Capsid Proteins; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000276:immunology* / Q000473:pathology; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D003432:Cross-Linking Reagents; D004705:Endocytosis; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D011991:Receptors, Virus / Q000037:antagonists & inhibitors / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D000914:Antibodies, Viral; D000951:Antigens, Neoplasm; D000956:Antigens, Viral; C568771:CLMP protein, human; C568772:CLMP protein, mouse; D036022:Capsid Proteins; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D003432:Cross-Linking Reagents; D007140:Immunoglobulin Fab Fragments; D011991:Receptors, Virus; C018315:hexon capsid protein, Adenovirus