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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.


Characterization of n-GO derivatives. (A) FT-IR measurements ofn-GO-alkyne before and after deuteration; (B) TGA graphs; (C) FT-IRmeasurements of n-GO-alkyne before and after click reaction. (D) SDS-PAGEgel electrophoresis under nonreducing (right) and reducing (left)conditions of anti-MUC1 IgG (1) and n-GO-PEG-MUC1 (2).
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fig1: Characterization of n-GO derivatives. (A) FT-IR measurements ofn-GO-alkyne before and after deuteration; (B) TGA graphs; (C) FT-IRmeasurements of n-GO-alkyne before and after click reaction. (D) SDS-PAGEgel electrophoresis under nonreducing (right) and reducing (left)conditions of anti-MUC1 IgG (1) and n-GO-PEG-MUC1 (2).

Mentions: FT-IR spectroscopy was used to confirm that the alkynegroup wasintroduced on the GO. However, the alkyne signal at 3292 cm–1 was masked by the O–H band from GO. For this reason deuteriumconversion was used to assess the presence of alkyne groups introducedon GO (Figure 1A) asthe new C–D band displays a stronger signal at 2565 cm–1 compared with the initial C–H band. The terminalhydrogen was replaced by deuterium following a mild and efficientprotocol described by Bew et al.10 (Figure S2A). The FT-IR spectrum (Figure S2A, top panel) showed the appearanceof a peak at 2565 cm–1 due to the vibration of theC–D bond, proving the conversion of propargyl alcohol to 2H-propargyl alcohol. Figure S2A shows the GO-alkyne spectrum with a new band appearing at 2565 cm–1, after deuteration that corresponds to the vibrationof the bond C–D. The O–H of GO may also be convertedto O–D, which overlaps with the C–D bond. To eliminatethis possibility, we subjected GO (lacking alkyne) to deuterationreaction conditions. No band was detected at 2565 cm–1 (Figure S2B), suggesting that the previouslyobserved peak at 2565 cm–1 was due to the deuteriumconversion.


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)

Characterization of n-GO derivatives. (A) FT-IR measurements ofn-GO-alkyne before and after deuteration; (B) TGA graphs; (C) FT-IRmeasurements of n-GO-alkyne before and after click reaction. (D) SDS-PAGEgel electrophoresis under nonreducing (right) and reducing (left)conditions of anti-MUC1 IgG (1) and n-GO-PEG-MUC1 (2).
© Copyright Policy
Related In: Results  -  Collection

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

fig1: Characterization of n-GO derivatives. (A) FT-IR measurements ofn-GO-alkyne before and after deuteration; (B) TGA graphs; (C) FT-IRmeasurements of n-GO-alkyne before and after click reaction. (D) SDS-PAGEgel electrophoresis under nonreducing (right) and reducing (left)conditions of anti-MUC1 IgG (1) and n-GO-PEG-MUC1 (2).
Mentions: FT-IR spectroscopy was used to confirm that the alkynegroup wasintroduced on the GO. However, the alkyne signal at 3292 cm–1 was masked by the O–H band from GO. For this reason deuteriumconversion was used to assess the presence of alkyne groups introducedon GO (Figure 1A) asthe new C–D band displays a stronger signal at 2565 cm–1 compared with the initial C–H band. The terminalhydrogen was replaced by deuterium following a mild and efficientprotocol described by Bew et al.10 (Figure S2A). The FT-IR spectrum (Figure S2A, top panel) showed the appearanceof a peak at 2565 cm–1 due to the vibration of theC–D bond, proving the conversion of propargyl alcohol to 2H-propargyl alcohol. Figure S2A shows the GO-alkyne spectrum with a new band appearing at 2565 cm–1, after deuteration that corresponds to the vibrationof the bond C–D. The O–H of GO may also be convertedto O–D, which overlaps with the C–D bond. To eliminatethis possibility, we subjected GO (lacking alkyne) to deuterationreaction conditions. No band was detected at 2565 cm–1 (Figure S2B), suggesting that the previouslyobserved peak at 2565 cm–1 was due to the deuteriumconversion.

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.