For Tf conjugation, 0.2 M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) solution was carefully stirred with Tf (5 mg/mL) and activated the Tf amine group for 2 hours. An added NGO-complex PEI solution then stood at 4°C overnight. Finally, reagents were removed by dialysis for 8 hours. Tf-NGO@Pt was analyzed with atomic-force microscopy (Bioscope Catalyst NanoScope V), transmission electron microscopy (Hitachi H-7650) with 80 kV accelerating voltage, and scanning electron microscopy (EX-250; Horiba). A Zetasizer Nano ZS particle analyzer (Malvern Instruments, Malvern, UK) was used to measured size distribution and ζ-potential of nanoparticles. Chemical construction of the nanosystem was characterized by ultraviolet (UV)-visible spectroscopy (Carry 5000) and Fourier-transform infrared (FTIR) spectroscopy (Equinox 55). A bicinchoninic acid kit (Pierce) was used to measure the concentration of Tf conjugate in the nanosystem. Pt content was determined as previously described by inductively coupled plasma atomic emission spectroscopy (ICP-AES).12 (link),29 (link),30 (link)
Protocol detail
Tf-NGO@Pt Nanosystem Characterization
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Amine
Atomic force microscopy
Bicinchoninic acid
Carbodiimide
Dialysis
Fourier transform infrared spectroscopy
Inductively coupled plasma atomic emission spectroscopy
Scanning electron microscopy
Spectroscopy
Transmission electron microscopy
Corresponding Organization :
Other organizations : Guangdong Provincial Hospital of Traditional Chinese Medicine, Jinan University
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Variable analysis
independent variables
- EDC concentration (0.2 M)
- Tf concentration (5 mg/mL)
- NGO-complex PEI solution
- Reaction duration (2 hours for EDC-Tf activation, overnight for NGO-complex PEI addition)
- Size distribution and zeta potential of Tf-NGO@Pt nanoparticles
- Chemical construction of the nanosystem (characterized by UV-visible and FTIR spectroscopy)
- Concentration of Tf conjugate in the nanosystem
- Pt content in the nanosystem
- Dialysis duration (8 hours) to remove reagents
- Analytical techniques used (atomic-force microscopy, transmission electron microscopy, scanning electron microscopy, Zetasizer Nano ZS, UV-visible spectroscopy, FTIR spectroscopy, bicinchoninic acid kit, inductively coupled plasma atomic emission spectroscopy)
- None specified
- None specified
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