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Genetically engineering encapsulin protein cage nanoparticle as a SCC-7 cell targeting optical nanoprobe.

Moon H, Lee J, Kim H, Heo S, Min J, Kang S - Biomater Res (2014)

Bottom Line: Encapsulin protein cage nanoparticle is used to develop a cell-specific targeting optical nanoprobe.FcBPs are genetically inserted and successfully displayed on the surface of encapsulin to form FcBP-encapsulin.Encapsulin protein cage nanoparticle is robust enough to maintain their structure at high temperature and easily acquires multifunctions on demand through the combination of genetic and chemical modifications.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 689-798 South Korea.

ABSTRACT

Background: Protein cage nanoparticles are promising nanoplatform candidates for efficient delivery systems of diagnostics and/or therapeutics because of their uniform size and structure as well as high biocompatibility and biodegradability. Encapsulin protein cage nanoparticle is used to develop a cell-specific targeting optical nanoprobe.

Results: FcBPs are genetically inserted and successfully displayed on the surface of encapsulin to form FcBP-encapsulin. Selectively binding of FcBP-encapsulin to SCC-7 is visualized with fluorescent microscopy.

Conclusions: Encapsulin protein cage nanoparticle is robust enough to maintain their structure at high temperature and easily acquires multifunctions on demand through the combination of genetic and chemical modifications.

No MeSH data available.


Fluorescent microscopic images. Fluorescent microscopic images of HepG2 cells (A,D) treated with f-encapsulin (A,B) and fFcBP-encapsulin (C,D). Fluorescent microscopic images of MDA-MB-231 cells (E,H) treated with f-encapsulin (E,F) and fFcBP-encapsulin (G,H). Fluorescent microscopic images of KB cells (I,L) treated with f-encapsulin (I,J) and fFcBP-encapsulin (K,L). DAPI (A, C, E, G, I, K) and fluorescein (B, D, F, H, J, L) were represented.
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Fig4: Fluorescent microscopic images. Fluorescent microscopic images of HepG2 cells (A,D) treated with f-encapsulin (A,B) and fFcBP-encapsulin (C,D). Fluorescent microscopic images of MDA-MB-231 cells (E,H) treated with f-encapsulin (E,F) and fFcBP-encapsulin (G,H). Fluorescent microscopic images of KB cells (I,L) treated with f-encapsulin (I,J) and fFcBP-encapsulin (K,L). DAPI (A, C, E, G, I, K) and fluorescein (B, D, F, H, J, L) were represented.

Mentions: To determine whether FcBP-encapsulin can bind a particular cell selectively, we treated a variety of cell lines with fFcBP-encapsulin and visualized them with fluorescent microscopy. F5M-conjugated encapsulin (f-encapsulin) was also treated in parallel as control. We first tried SCC-7 cells which overexpress a cell surface glycoprotein CD44 involved in cell-cell interactions, cell adhesion and migration. While f-encapsulin did not bind to SCC-7 cells at all (Figure 3A-C), fFcBP-encapsulin bound to SCC-7 cells (Figure 3D-F). Fluorescence images of SCC-7 treated with fFcBP-encapsulin showed cytosolic accumulation of fFcBP-encapsulin (Figure 3E, F). These data suggest that FcBP-encapsulin recognizes surface markers of SCC-7 cells, selectively binds to them, and is internalized. Furthermore, the pretreatment of SCC-7 cells with anti-CD44 antibody completely blocked the binding of FcBP-encapsulin (data not shown). We then chose Hela cells and treated them with same way of SCC-7 cells. We did not observe any binding of neither fFcBP-encapsulin nor f-encapsulin to Hela cells (Figure 3G-L). To investigate cell selectivity of FcBP-encapsulin further, we prepared HepG2 hepatocyte cells, MDA-MB-231 breast cancer cells, and KB epithelial cells additionally and treated them with fEcBP-encapsulin or f-encapsulin. None of cell lines we prepared showed the evidence of specific binding of either fEcBP-encapsulin or f-encapsulin (Figure 4). Although through studies about surface molecules that FcBP-encapsulin binds to should be done, these data demonstrate that FcBP-encapsulin selectively recognizes and binds to SCC-7 cells and has a potential to be used as an efficient molecular imaging probe in vitro. FcBP-encapsulin has a large internal cavity (20 nm in diameter) and it can be used for encapsulating chemicals, nanomaterials, and proteins. Thus, encapsulin has a potential to be used as a modular template for developing a versatile, multifunctional theranostic system, which has specific cell targeting ligands and diagnostic and therapeutic reagents simultaneouslyFigure 3


Genetically engineering encapsulin protein cage nanoparticle as a SCC-7 cell targeting optical nanoprobe.

Moon H, Lee J, Kim H, Heo S, Min J, Kang S - Biomater Res (2014)

Fluorescent microscopic images. Fluorescent microscopic images of HepG2 cells (A,D) treated with f-encapsulin (A,B) and fFcBP-encapsulin (C,D). Fluorescent microscopic images of MDA-MB-231 cells (E,H) treated with f-encapsulin (E,F) and fFcBP-encapsulin (G,H). Fluorescent microscopic images of KB cells (I,L) treated with f-encapsulin (I,J) and fFcBP-encapsulin (K,L). DAPI (A, C, E, G, I, K) and fluorescein (B, D, F, H, J, L) were represented.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Fig4: Fluorescent microscopic images. Fluorescent microscopic images of HepG2 cells (A,D) treated with f-encapsulin (A,B) and fFcBP-encapsulin (C,D). Fluorescent microscopic images of MDA-MB-231 cells (E,H) treated with f-encapsulin (E,F) and fFcBP-encapsulin (G,H). Fluorescent microscopic images of KB cells (I,L) treated with f-encapsulin (I,J) and fFcBP-encapsulin (K,L). DAPI (A, C, E, G, I, K) and fluorescein (B, D, F, H, J, L) were represented.
Mentions: To determine whether FcBP-encapsulin can bind a particular cell selectively, we treated a variety of cell lines with fFcBP-encapsulin and visualized them with fluorescent microscopy. F5M-conjugated encapsulin (f-encapsulin) was also treated in parallel as control. We first tried SCC-7 cells which overexpress a cell surface glycoprotein CD44 involved in cell-cell interactions, cell adhesion and migration. While f-encapsulin did not bind to SCC-7 cells at all (Figure 3A-C), fFcBP-encapsulin bound to SCC-7 cells (Figure 3D-F). Fluorescence images of SCC-7 treated with fFcBP-encapsulin showed cytosolic accumulation of fFcBP-encapsulin (Figure 3E, F). These data suggest that FcBP-encapsulin recognizes surface markers of SCC-7 cells, selectively binds to them, and is internalized. Furthermore, the pretreatment of SCC-7 cells with anti-CD44 antibody completely blocked the binding of FcBP-encapsulin (data not shown). We then chose Hela cells and treated them with same way of SCC-7 cells. We did not observe any binding of neither fFcBP-encapsulin nor f-encapsulin to Hela cells (Figure 3G-L). To investigate cell selectivity of FcBP-encapsulin further, we prepared HepG2 hepatocyte cells, MDA-MB-231 breast cancer cells, and KB epithelial cells additionally and treated them with fEcBP-encapsulin or f-encapsulin. None of cell lines we prepared showed the evidence of specific binding of either fEcBP-encapsulin or f-encapsulin (Figure 4). Although through studies about surface molecules that FcBP-encapsulin binds to should be done, these data demonstrate that FcBP-encapsulin selectively recognizes and binds to SCC-7 cells and has a potential to be used as an efficient molecular imaging probe in vitro. FcBP-encapsulin has a large internal cavity (20 nm in diameter) and it can be used for encapsulating chemicals, nanomaterials, and proteins. Thus, encapsulin has a potential to be used as a modular template for developing a versatile, multifunctional theranostic system, which has specific cell targeting ligands and diagnostic and therapeutic reagents simultaneouslyFigure 3

Bottom Line: Encapsulin protein cage nanoparticle is used to develop a cell-specific targeting optical nanoprobe.FcBPs are genetically inserted and successfully displayed on the surface of encapsulin to form FcBP-encapsulin.Encapsulin protein cage nanoparticle is robust enough to maintain their structure at high temperature and easily acquires multifunctions on demand through the combination of genetic and chemical modifications.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 689-798 South Korea.

ABSTRACT

Background: Protein cage nanoparticles are promising nanoplatform candidates for efficient delivery systems of diagnostics and/or therapeutics because of their uniform size and structure as well as high biocompatibility and biodegradability. Encapsulin protein cage nanoparticle is used to develop a cell-specific targeting optical nanoprobe.

Results: FcBPs are genetically inserted and successfully displayed on the surface of encapsulin to form FcBP-encapsulin. Selectively binding of FcBP-encapsulin to SCC-7 is visualized with fluorescent microscopy.

Conclusions: Encapsulin protein cage nanoparticle is robust enough to maintain their structure at high temperature and easily acquires multifunctions on demand through the combination of genetic and chemical modifications.

No MeSH data available.