Investigation of MiRGD peptide and Graphene Quantum Dots nano-carrier for targeted delivery of Aflibercept to retinoblastoma cancer cells.
Naeimeh Bayatkhani1 , Zahra-Soheila Soheili1 *, Saman Hosseinkhani2 , Hamid Ahmadieh3 , Somayeh Piroozmand1 , Hamid Latifi-Navid4 , Sina Goli Garmestani1 , Shahram Samiei5
- Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran 1497716316, Iran
- Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran and Department of Nanobiotechnology, Faculty of Biological Sciences,Tarbiat Modares University, Tehran, Iran
- Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
- Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran 1497716316, Iran and Electrophysiology Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran 1416634793, Iran and School of Biological Sciences, Institute for Research in Fundamental Sciences (IPM), Tehran, Iran
- Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
Abstract: Although Retinoblastoma is known as a curable cancer, untreated one can be fatal and even with treatment, advanced tumors can limit globe salvage. Therefore, there is a necessity for designing a new drug delivery system. A novel nano-carrier with MiRGD peptides and graphene quantum dots (GQDs) has been developed due to the structural differences between cancerous and normal cells. The iRGD motif penetrates cancerous tissues, while the other motifs deliver both hydrophobic and hydrophilic drugs. This peptide is suitable for delivering an anti-VEGF drug to prevent angiogenesis activation. GQDs can aid in efficient biological tracking as non-toxic particles and enhance drug binding to peptides.
Methods: To purify the MiRGD peptide, Ecoli BL21 was cultured in a 2XYT medium containing kanamycin antibiotic and IPTG as inducer of protein expression. After using a lysis buffer, the MiRGD peptide was purified by Ni-NTA column chromatography under a urea gradient. Briefly, the impurities were removed by washing buffers. The purity of the peptide was examined by SDS-PAGE. Purified peptide was desalted by dialysis against PBS buffer.
The drug solution will be mixed with varying concentrations of the purified peptide separately. GQDs were synthesized hydrothermally by dissolving citric acid and urea in water. Then ethanol was added into the solution and the final product was collected by centrifuging. The Solution was dried and dissolved into deionized water. For the next experiments GQDs-MiRGD- Aflibercept drug complexes will be prepared.
Results: The MiRGD peptide band was observed on 15% Tris – glycine SDS-PAGE gel. UV/Vis absorption spectroscopy will be performed for synthesized GQDs to display its two peaks at 234 nm and 337 nm. The bands related to surface functional groups present on GQDs will be detected by FTIR. The fluorescence spectrum of the synthesized GQDs will be investigated at different excitation wavelengths. Correct admissible formation of GQDs-MiRGD- Aflibercept drug complex will be investigated later.
Conclusion: To create a new nano-carrier for targeted drug delivery, MiRGD peptide and GQDs were synthesized and characterized. For the next step the complex of MiRGD, drug and GQDs are going to be assembled and analyzed.
