Enhanced cellular uptake of virus-like particles through immobilization on a sialic acid-displaying solid surface.
Herein, we present the efficient cellular uptake of immobilized virus-like particles (VLPs) made of recombinant JC virus capsid proteins. VLPs expressed in Escherichia coli were labeled with fluorescein isothiocyanate (FITC). We compared two approaches for the cellular uptake of the FITC-VLPs. In the first approach, FITC-VLPs were immobilized on a polystyrene substrate, and then NIH3T3 cells were cultured on the same substrate. In the second approach, cells were cultured on a polystyrene substrate, and FITC-VLPs were then added to the cell culture medium. Flow cytometric analysis and confocal laser microscopic observation revealed that immobilized VLPs were incorporated into the cells with higher efficiency than were the diffusive VLPs suspended in solution. The cellular uptake of VLPs on the substrate was increased in a VLP density-dependent manner. As a control, disassembling VLPs to form VP1 pentamers abolished incorporation into the cells. Displaying sialic acid on the substrate enhanced VLP density through the specific affinities between the VLPs and sialic acid, resulting in efficient incorporation into the seeded cells. These techniques can be applied to the development of novel drug delivery systems and cell microarrays not only of nucleic acids but also of small molecules and proteins through their encapsulation in VLPs.
Ohtake
Noriko
N
Graduate School of Science, Hokkaido University N21W10, Sapporo 001-0021, Japan.
Niikura
Kenichi
K
Suzuki
Tadaki
T
Nagakawa
Keita
K
Sawa
Hirofumi
H
Ijiro
Kuniharu
K
D000818:Animals; D005434:Flow Cytometry; D051379:Mice; D018613:Microscopy, Confocal; D019158:N-Acetylneuraminic Acid / Q000737:chemistry*; D041681:NIH 3T3 Cells; D020349:Surface Plasmon Resonance; D014771:Virion / Q000378:metabolism*
D019158:N-Acetylneuraminic Acid