PubMed | Childrens Hospital Oakland Research Institute, Northwestern University and a Lypro Biosciences Inc.
Type: Journal Article | Journal: Biochemistry and cell biology = Biochimie et biologie cellulaire | Year: 2015
A fusion protein comprising an -CD20 single chain variable fragment (scFv) antibody, a spacer peptide, and human apolipoprotein (apo) A-I was constructed and expressed in Escherichia coli. The lipid interaction properties intrinsic to apoA-I as well as the antigen recognition properties of the scFv were retained by the chimera. scFvapoA-I was formulated into nanoscale reconstituted high-density lipoprotein particles (termed nanodisks; ND) and incubated with cultured cells. -CD20 scFvapoA-I ND bound to CD20-positive non-Hodgkins lymphoma (NHL) cells (Ramos and Granta) but not to CD20-negative T lymphocytes (i.e., Jurkat). Binding to NHL cells was partially inhibited by pre-incubation with rituximab, a monoclonal antibody directed against CD20. Confocal fluorescence microscopy analysis of Granta cells following incubation with -CD20 scFvapoA-I ND formulated with the intrinsically fluorescent hydrophobic polyphenol, curcumin, revealed -CD20 scFvapoA-I localizes to the cell surface, while curcumin off-loads and gains entry to the cell. Compared to control incubations, viability of cultured NHL cells was decreased upon incubation with -CD20 scFvapoA-I ND harboring curcumin. Thus, formulation of curcumin ND with -CD20 scFvapoA-I as the scaffold component confers cell targeting and enhanced bioactive agent delivery, providing a strategy to minimize toxicity associated with chemotherapeutic agents.