Selective internalization of self-assembled artificial oil bodies by HER2/neu-positive cells.
A novel delivery carrier was developed using artificial oil bodies (AOBs). Plant seed oil bodies (OBs) consist of a triacylglycerol matrix surrounded by a monolayer of phospholipids embedded with the storage protein oleosin (Ole). Ole consists of a central hydrophobic domain with two amphiphatic arms that extrude from the surface of OBs. In this study, a bivalent anti-HER2/neu affibody domain (ZH2) was fused with Ole at the C terminus. After overproduction in Escherichia coli, the fusion protein (Ole-ZH2) was recovered to assemble AOBs. The size of self-assembled AOBs was tailored by varying the oil/Ole-ZH2 ratio and pH to reach a nanoscale. Upon co-incubation with tumor cells, the nanoscale AOBs encapsulated with a hydrophobic fluorescence dye were selectively internalized by HER2/neu-overexpressing cells and displayed biocompatibility with the cells. In addition, the ZH2-mediated endosomal entry of AOBs occurred in a time- and AOB dose-dependent manner. The internalization efficiency was as high as 90%. The internalized AOBs disintegrated at the non-permissive pH (e.g. in acidic endosomes) and the cargo dye was released. Results of in vitro study revealed a sustained and prolonged release profile. Taken together, our findings indicate the potential of AOBs as a delivery carrier.
Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung, Taiwan. cjchiang@mail.cmu.edu.tw
Chung-Jen
CJ
Chiang
Li-Jen
LJ
Lin
Che-Chin
CC
Lin
Chih-Hsiang
CH
Chang
Yun-Peng
YP
Chao
D058045:Artificial Cells / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D045744:Cell Line, Tumor; D002470:Cell Survival; D003001:Cloning, Molecular; D004355:Drug Stability; D004705:Endocytosis; D005434:Flow Cytometry; D006801:Humans; D008422:Materials Testing; D008856:Microscopy, Fluorescence; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*; D012639:Seeds / Q000737:chemistry; D014280:Triglycerides / Q000737:chemistry / Q000378:metabolism
D010743:Phospholipids; D010940:Plant Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; D014280:Triglycerides