1. 11 SYNTHETIC PEPTIDE MEDIATED IMMUNOCONJUGATION OF QUANTUM DOTS Libor Janů 1, Maja Stanisavljević 1, Markéta Ryvolová 1,2, Jaromír Hubálek 2, Vojtěch Adam 1,2, René Kizek 1,2 Supervisor: Prof. Ing. René Kizek, PhD. 1 Department of Chemistry and Biochemistry, Faculty of agronomy, Mendel University 2 Central European Institute of Technology, Brno University of Technology

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

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

4. 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. 5 Results Fig.1: HPLC chromatogram of crude peptide HWRGWVC +tBu HWRGWVC

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

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

8. 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. 99 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 from IGA AF Mendelu, IP 10/2012.

10. 10 Thank you for your attention!