Limits...
Solvent-Free Click-Mechanochemistry for the Preparation of Cancer Cell Targeting Graphene Oxide.

Rubio N, Mei KC, Klippstein R, Costa PM, Hodgins N, Wang JT, Festy F, Abbate V, Hider RC, Chan KL, Al-Jamal KT - ACS Appl Mater Interfaces (2015)

Bottom Line: Polyethylene glycol-functionalized nanographene oxide (PEGylated n-GO) was synthesized from alkyne-modified n-GO, using solvent-free click-mechanochemistry, i.e., copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC).The modified n-GO was subsequently conjugated to a mucin 1 receptor immunoglobulin G antibody (anti-MUC1 IgG) via thiol-ene coupling reaction. n-GO derivatives were characterized with Fourier-transformed infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), Bradford assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and atomic force microscopy (AFM).Cell targeting was confirmed in vitro in MDA-MB-231 cells, either expressing or lacking MUC1 receptors, using flow cytometry, confocal laser scanning microscopy (CLSM) and multiphoton (MP) fluorescence microscopy.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pharmaceutical Science, King's College London , Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, United Kingdom.

ABSTRACT
Polyethylene glycol-functionalized nanographene oxide (PEGylated n-GO) was synthesized from alkyne-modified n-GO, using solvent-free click-mechanochemistry, i.e., copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC). The modified n-GO was subsequently conjugated to a mucin 1 receptor immunoglobulin G antibody (anti-MUC1 IgG) via thiol-ene coupling reaction. n-GO derivatives were characterized with Fourier-transformed infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), Bradford assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and atomic force microscopy (AFM). Cell targeting was confirmed in vitro in MDA-MB-231 cells, either expressing or lacking MUC1 receptors, using flow cytometry, confocal laser scanning microscopy (CLSM) and multiphoton (MP) fluorescence microscopy. Biocompatibility was assessed using the modified lactate dehydrongenase (mLDH) assay.

No MeSH data available.


Related in: MedlinePlus

Intracellularuptake of n-GO-PEG-MUC1 in MUC1+ and MUC1- humanbreast cancer cells in vitro. (A) Flow cytometric profiles of MUC1+and MUC1- cell lines treated with antiMUC1-IgG or GO-PEG-MUC1 for1 h, 3 or 24 h. Cellular uptake was assessed by analyzing the shiftin mean fluorescence intensity (MFI). The percentage of positive cellsfor MUC1− and MUC1+ cells are shown in (B) and (C). Signalswere collected using FL2 detector.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4559840&req=5

fig3: Intracellularuptake of n-GO-PEG-MUC1 in MUC1+ and MUC1- humanbreast cancer cells in vitro. (A) Flow cytometric profiles of MUC1+and MUC1- cell lines treated with antiMUC1-IgG or GO-PEG-MUC1 for1 h, 3 or 24 h. Cellular uptake was assessed by analyzing the shiftin mean fluorescence intensity (MFI). The percentage of positive cellsfor MUC1− and MUC1+ cells are shown in (B) and (C). Signalswere collected using FL2 detector.

Mentions: The uptake of n-GO-PEG-MUC1 in MUC1+ cells was also assessedbyflow cytometry and the extent of uptake was expressed as mean fluorescenceintensity (MFI) (Figure 3). Cells were treated for 1, 3, and 24 h with anti-MUC1 IgG or n-GO-PEG-MUC1at concentrations equivalent to 2.5 μg/mL anti-MUC1 IgG. n-GO-PEGwas used as a control. At 1 h, comparable results were obtained forboth anti-MUC1 IgG and n-GO-PEG-MUC1 with uptake only being observedin MUC1+ cells. At 3 h, an increase in MFI was found in MUC1–cells treated with n-GO-PEG-MUC1. This increase in MFI, however, wassignificantly lower than that observed in MUC1+ cells. At 24 h, MUC1+cells treated with n-GO-PEG-MUC1 showed significantly higher MFI valuescompared to all treatment groups (Figure 3A), agreeing with CLSM and MP results.


Solvent-Free Click-Mechanochemistry for the Preparation of Cancer Cell Targeting Graphene Oxide.

Rubio N, Mei KC, Klippstein R, Costa PM, Hodgins N, Wang JT, Festy F, Abbate V, Hider RC, Chan KL, Al-Jamal KT - ACS Appl Mater Interfaces (2015)

Intracellularuptake of n-GO-PEG-MUC1 in MUC1+ and MUC1- humanbreast cancer cells in vitro. (A) Flow cytometric profiles of MUC1+and MUC1- cell lines treated with antiMUC1-IgG or GO-PEG-MUC1 for1 h, 3 or 24 h. Cellular uptake was assessed by analyzing the shiftin mean fluorescence intensity (MFI). The percentage of positive cellsfor MUC1− and MUC1+ cells are shown in (B) and (C). Signalswere collected using FL2 detector.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4559840&req=5

fig3: Intracellularuptake of n-GO-PEG-MUC1 in MUC1+ and MUC1- humanbreast cancer cells in vitro. (A) Flow cytometric profiles of MUC1+and MUC1- cell lines treated with antiMUC1-IgG or GO-PEG-MUC1 for1 h, 3 or 24 h. Cellular uptake was assessed by analyzing the shiftin mean fluorescence intensity (MFI). The percentage of positive cellsfor MUC1− and MUC1+ cells are shown in (B) and (C). Signalswere collected using FL2 detector.
Mentions: The uptake of n-GO-PEG-MUC1 in MUC1+ cells was also assessedbyflow cytometry and the extent of uptake was expressed as mean fluorescenceintensity (MFI) (Figure 3). Cells were treated for 1, 3, and 24 h with anti-MUC1 IgG or n-GO-PEG-MUC1at concentrations equivalent to 2.5 μg/mL anti-MUC1 IgG. n-GO-PEGwas used as a control. At 1 h, comparable results were obtained forboth anti-MUC1 IgG and n-GO-PEG-MUC1 with uptake only being observedin MUC1+ cells. At 3 h, an increase in MFI was found in MUC1–cells treated with n-GO-PEG-MUC1. This increase in MFI, however, wassignificantly lower than that observed in MUC1+ cells. At 24 h, MUC1+cells treated with n-GO-PEG-MUC1 showed significantly higher MFI valuescompared to all treatment groups (Figure 3A), agreeing with CLSM and MP results.

Bottom Line: Polyethylene glycol-functionalized nanographene oxide (PEGylated n-GO) was synthesized from alkyne-modified n-GO, using solvent-free click-mechanochemistry, i.e., copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC).The modified n-GO was subsequently conjugated to a mucin 1 receptor immunoglobulin G antibody (anti-MUC1 IgG) via thiol-ene coupling reaction. n-GO derivatives were characterized with Fourier-transformed infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), Bradford assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and atomic force microscopy (AFM).Cell targeting was confirmed in vitro in MDA-MB-231 cells, either expressing or lacking MUC1 receptors, using flow cytometry, confocal laser scanning microscopy (CLSM) and multiphoton (MP) fluorescence microscopy.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pharmaceutical Science, King's College London , Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, United Kingdom.

ABSTRACT
Polyethylene glycol-functionalized nanographene oxide (PEGylated n-GO) was synthesized from alkyne-modified n-GO, using solvent-free click-mechanochemistry, i.e., copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC). The modified n-GO was subsequently conjugated to a mucin 1 receptor immunoglobulin G antibody (anti-MUC1 IgG) via thiol-ene coupling reaction. n-GO derivatives were characterized with Fourier-transformed infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), Bradford assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and atomic force microscopy (AFM). Cell targeting was confirmed in vitro in MDA-MB-231 cells, either expressing or lacking MUC1 receptors, using flow cytometry, confocal laser scanning microscopy (CLSM) and multiphoton (MP) fluorescence microscopy. Biocompatibility was assessed using the modified lactate dehydrongenase (mLDH) assay.

No MeSH data available.


Related in: MedlinePlus