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SYNTHETIC PEPTIDE MEDIATED IMMUNOCONJUGATION OF QUANTUM DOTS

SYNTHETIC PEPTIDE MEDIATED IMMUNOCONJUGATION OF QUANTUM DOTS. Libor Janů 1 , Maja Stanisavljevi ć 1 , Markéta Ryvolová 1 ,2 , Jaromír Hubálek 2 , Vojtěch Adam 1 ,2 , René Kizek 1 ,2 Supervisor: Prof. Ing. René Kizek, PhD.

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SYNTHETIC PEPTIDE MEDIATED IMMUNOCONJUGATION OF QUANTUM DOTS

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  1. SYNTHETIC PEPTIDE MEDIATED IMMUNOCONJUGATION OF QUANTUM DOTS Libor Janů1, Maja Stanisavljević1, Markéta Ryvolová1,2, Jaromír Hubálek2, Vojtěch Adam1,2, René Kizek1,2 Supervisor: Prof. Ing. René Kizek, PhD. 1Department of Chemistry and Biochemistry,Faculty of agronomy,Mendel University 2Central European Institute of Technology, Brno University of Technology 1

  2. ANTIBODY NH2 LINKER HOOC QD Introduction • Immunochemistry – antigen-antibody reaction • Quantum dot labeled antibody • Standard conjugation chemistry (EDC, Sulfo-SMCC, ...) → non-specific reaction, lower QDs stability, multistep process

  3. ANTIBODY Protein A SBP QD Introduction • Alternative coupling strategy – electrostatic forces, polyhistidine tag, SBP + protein-protein interaction.

  4. Material and methods Synthesis of peptide HWRGWVC coated CdTe quantum dots • Microvawe synthesis: 180°C, 800W, 20 minutes Conjugation of peptide-quantum dots to the HIgG • Peptide HWRGWVC bind the Fc region of HIgG • 13 µl of HIgG was added to 100 µl of quantum dots

  5. Results Fig.1: HPLC chromatogram of crude peptide HWRGWVC HWRGWVC +tBu

  6. Results Fig.2: Emmision spectra of HWR quantum dots Excitation 250 nm Excitation 310 nm Excitation 480 nm

  7. Results Fig.3:Electroforeogram of crude QD-HWR-IgG 100x diluted sample HWR-QD and negative control HWR-QD-HIgG

  8. Conclusion • We successfully performed a peptide mediated quantum dot–HIgG coupling • Peptide coated quantum dots should bind the Fc region of HIgG • Relative small size of final complex • Preparation of immuno-quantum dots is based on simple mixing of quantum dots and HIgG • Solubility of peptide in quantum dots reaction mixture – next improvement of the process

  9. Acknowledgements • prof. Ing. René Kizek, PhD. • Ing. Markéta Ryvolová, PhD. • Maja Stanisavljevic • Mgr. Natalia Cernei • Ing. Pavlína Šobrová • doc. RNDr. Pavel Kopel, Ph.D. • This study was supported fromIGA AF Mendelu, IP 10/2012. 9

  10. Thank you for your attention! 10

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