Design strategies
that lead to a more focused in vivo
delivery
of functionalized nanoparticles (NPs) and their cargo can potentially
maximize their therapeutic efficiency while reducing systemic effects,
broadening their clinical applications. Here, we report the development
of a noncovalent labeling approach where immunoglobulin G (IgG)-decorated
NPs can be directed to a cancer cell using a simple, linear bispecific
protein adaptor, termed MFE23-ZZ. MFE23-ZZ was created by fusing a
single-chain fragment variable domain, termed MFE23, recognizing carcinoembryonic
antigen (CEA) expressed on tumor cells, to a small protein ZZ module,
which binds to the Fc fragment of IgG. As a proof of concept, monoclonal
antibodies (mAbs) were generated against a NP coat protein, namely,
gas vesicle protein A (GvpA) of Halobacterium salinarum gas vesicles (GVs). The surface of each GV was therapeutically derivatized
with the photoreactive agent chlorin e6 (Ce6GVs) and anti-GvpA mAbs were subsequently bound to GvpA
on the surface of each Ce6GV. The bispecific ligand
MFE23-ZZ was then bound to mAb-decorated Ce6GVs via
their Fc domain, resulting in a noncovalent tripartite complex, namely,
MFE23.ZZ-2B10–Ce6GV. This complex enhanced
the intracellular uptake of Ce6GVs into human CEA-expressing
murine MC38 colon carcinoma cells (MC38.CEA) relative to the CEA-negative
parental cell line MC38 in vitro, making them more sensitive to light-induced
cell killing. These results suggest that the surface of NP can be
rapidly and noncovalently functionalized to target tumor-associated
antigen-expressing tumor cells using simple bispecific linkers and
any IgG-labeled cargo. This noncovalent approach is readily applicable
to other types of functionalized NPs.