U. Wojda et Jl. Miller, Targeted transfer of polyethylenimine-avidin-DNA bioconjugates to hematopoietic cells using biotinylated monoclonal antibodies, J PHARM SCI, 89(5), 2000, pp. 674-681
Here we examine whether attachment of biotinylated antibodies to proteins o
n the cell surface increases the transfection efficiency of polyethylenimin
e-avidin-DNA bioconjugate gene transfer. Preliminary experiments were perfo
rmed to compare avidin endocytosis into cells incubated with biotinylated a
ntibodies. Antibody biotinylation resulted in the endocytosis of avidin-FIT
C into nearly 100% of cells compared with no detectable binding or entry in
to unbiotinylated cells. Gene transfer was accomplished with avidin conjuga
ted to polyethylenimine (PEI) at a molar ratio of 4:1 (PA4). Plasmid DNA en
coding the green fluorescent protein (GFP) gene was condensed on the PA4, a
nd transfection efficiencies were measured by flow cytometry as the percent
age of cells that fluoresced at levels greater than two standard deviations
above the negative control. Gene transfer efficiencies were compared among
K562, HEL, and Jurkat leukemia cell lines. Control transfections with DNA
alone or untargeted PEI-DNA resulted in less than or equal to 2% GFP positi
ve cells. Targeting PEI-avidin-DNA to antibody biotinylated cells increased
transfection efficiency several fold over untargeted PEI. For each cell ty
pe, the increase in transfection efficiency was not significantly different
among four biotinylated antibodies tested (antiCD55, antiCD59, antiCD71, a
nd antiCD98). These data suggest biotinylated antibodies may be useful for
targeting polyethylenimine-avidin mediated gene transfer. (C) 2000 Wiley-Li
ss, Inc.