Hücrelerin Hedeflenmesi ve Görüntülenmesi için Floresans Prob Olarak Kuantum Nokta Biyokonjugatlarının Hazırlanması

B urada, hedeflenmiş hücresel görüntüleme için suda çözünür kuantum noktaların QD biyokonjugasyonu ve uygulamaları gerçekleştirilmiştir. Kanser hücrelerine nanoyapıların hedeflenmesi için anti-c-ErbB2 antikoru QD’lara bağlanmıştır. QD’lar ve anti-c-ErbB2/QD biyokonjugatları floresans/UV-Görünür bölge spektroskopisi ve hidrodinamik boyut ölçümleri ile karakterize edilmiştir. Hücre canlılığına QD’lar ve antic-ErbB2/QD biyokonjugatlarının etkisi MTT denemeleri kullanılarak gerçekleştirilmiştir. Anti-c-ErbB2/QD biyokonjugatlarının hedefleme etkinliğini gözlemlemek için, floresans mikroskopi kullanılmıştır. Hücreleri görüntüleme ve iyi gerçekleştirilmiş hedeflemenin ispatı olarak anti-c-ErbB2 konjuge edilen QD’lar ile daha yüksek hücre afiniteleri elde edilmiştir

Anahtar Kelimeler:

Kuantum noktalar, biyokonjugasyon, anti-c-ErbB2, hücre hedefleme

Preparation of Quantum Dot Bioconjugates as Fluorescence Probe for Targeting and Visualization of Cells

Here, the bioconjugation and application of water-soluble quantum dots QDs for targeted cellular imaging were investigated. Human Anti-c-ErbB2 antibodies were bound to QDs for the targeting of these nanostructures towards the cancer cells. QDs and Anti-c-ErbB2/QD bioconjugates were characterized by fluorescence and UV–Vis spectroscopy and hydrodynamic sizing. The effect of QDs and Anti-c-ErbB2/QD bioconjugates on cell viability was evaluated using an MTT assay. To observe targeting efficiency of Anti-c- ErbB2/QD bioconjugates, fluorescence microscopy and was performed. Higher cell affinities were obtained by Anti-c-ErbB2-conjugated QDs which is an evidence that these are well-suited for targeting and imaging of cells.

Keywords:

Quantum dots, bioconjugation, anti-c-ErbB2, cell targeting,

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1. D. Odaci Demirkol, S. Timur, A sandwich-type assay based on quantum dot/aptamer bioconjugates for analysis of E. coli O157:H7 in microtiter plate format, Int. J. Polym. Mater., 65 (2016) 85-90.
2. M. Akin, R. Bongartz, J.G. Walter, D. Odaci Demirkol, F. Stahl, S. Timur, T. Scheper,PAMAM-functionalized water soluble quantum dots for cancer cell targeting, J. Mater. Chem. B, 22 (2012) 11529-11536.
3. D. Ag, L. Eral Dogan, R. Bongartz, M. Seleci, J.G. Walter, D. Odaci Demirkol, F. Stahl, S. Ozcelik, S. Timur, T. Scheper, Biofunctional quantum dots as fluorescence probe for cell specific targeting, Coll. Surf. B, 114 (2014) 96-103.
4. M. Kesik, B. Demir, F.B. Barlas, C. Geyik, S.C. Cevher, D. Odaci Demirkol, S. Timur, A. Cirpan, L. Toppare, Multi-functional fluorescent scaffolds as a multi-color probe: Design and application in targeted cell imaging, RSC Adv., 5 (2015) 83361-83367.
5. D. Goen Colak, I. Cianga, D. Odaci Demirkol, O. Kozgus, I. Medine, S. Sakarya, P. Unak, S. Timur, Y. Yagci, The synthesis and targeting of PPP-type copolymers to breast cancer cells: multifunctional platforms for imaging and diagnosis, J. Mater. Chem. B, 22 (2012) 9293-9300.
6. C. Geyik, E. Guler, Z.P. Gumus, F.B. Barlas, H. Akbulut, D. Odaci Demirkol, S. Timur, Y. Yagci, Bioconjugation and applications of amino functional fluorescence polymers, Macromol. Biosci., 17 (2017) 1600232.
7. E. Guler, H. Akbulut, C. Geyik, T. Yilmaz, Z.P. G u m u s , F.B. Barlas, R.E. Ahan, D. Odaci Demirkol, S. Yamada, T. Endo, S. Timur, Y. Yagci, Complex structured fluorescent polythiophene graft copolymer as a versatile tool for imaging, targeted delivery of paclitaxel, and radiotherapy, Biomacromolecules, 17 (2016) 2399–2408.
8. M. Geszke-Moritz, M. Moritz, Quantum dots as versatile probes in medical sciences: Synthesis, modification and properties, Mat. Sci. Eng. C- Mater., 33 (2013) 1008–1021.
9. S.J. Rosenthal, J.C. Chang, O. Kovtun, J.R. McBride, I.D. Tomlinson, Biocompatible quantum dots for biological applications, Chem. Biol., 18 (2011) 10-24.
10. Y. Ghasemi, P. Peymani, S. Afifi, Quantum dot: magic nanoparticle for imaging, detection and targeting, Acta Biomed., 80 (2009) 156-165.
11. T. R. Pisanic, Y. Zhang, T. H. Wang, Quantum dots in diagnostics and detection: principles and paradigms, Analyst, 139 (2014) 2968-2981.
12. C. M. Tyrakowski, P.T. Snee, A primer on the synthesis, water-solubilization, and functionalization of quantum dots, their use as biological sensing agents, and present status, Phys. Chem. Chem. Phys., 16 (2014) 837-855.
13. A.M. Smith, S. Dave, S. Nie, L. True, X. Gao, Multicolor quantum dots for molecular diagnostics of cancer, Expert Rev. Mol. Diagn., 6 (2006) 231-244.
14. D.J. Bharali, D.W. Lucey, H. Jayakumar, H.E. Pudavar, P.N. Prasad, Folate-receptor-mediated delivery of InP quantum dots for bioimaging using confocal and twophoton microscopy, J. Am. Chem. Soc., 127 (2005) 11364-11371.
15. Z. Li, P. Huang, J. Lin, R. He, B. Liu, X. Zhang, S. Yang, P. Xi, X. Zhang, Q. Ren, D. Cui, Arginine-glycine-aspartic acid-conjugated dendrimer-modified quantum dots for targeting and imaging melanoma, J. Nanosci. Nanotechno., 10 (2010) 1–9.
16. J. Ruan, H. Song, Q. Qian, C. Li, K. Wang, C. Bao, D. Cui, HER2 monoclonal antibody conjugated RNase-Aassociated CdTe quantum dots for targeted imaging and therapy of gastric cancer, Biomaterials, 33 (2012) 7093-7102.
17. B. Seven, M. Bourourou, K. Elouarzaki, J.F. Constant, C. Gondran, M. Holzinger,S. Cosnier, S. Timur, Impedimetric biosensor for cancer cell detection, Electrochem. Commun., 37 (2013) 36–39.
18. K.W. Kim, J. Song, J.S. Kee, Q. Liu, G.Q. Lo, M.K. Park, Label-free biosensor based on an electrical tracing-assisted silicon mirroring resonator with a low-cost broad band source, Biosens. Bioelectron., 46 (2013) 15–21.
19. K. Manzoor, S. Johny, D. Thomas, S. Setua, D. Menon, S. Nair, Bio-conjugated luminescent quantum dots of doped ZnS: a cyto-friendly system for targeted cancer imaging, Nanotechnology, 20 (2009) 65-102.
20. M. Geszke, M. Murias, L. Balan, G. Medjahdi, J. Korczynski, M. Moritz, J. Lulek, R. Schneider, Folic acid-conjugated core/shell ZnS:Mn/ZnS quantum dots as targeted probes for two photon fluorescence imaging of cancer cells, Acta Biomater., 7 (2010) 1327-1338.