1. Nanomaterial’s Application in Biotechnology Lei Sun Project manager Digital Health Group Intel Corp.

2. Outline Overview of nanomaterial’s application in biotech, especially in cancer diagnosis and treatment. Application Example I: Gold Nanoparticle’s application in DNA detection. Application example II: Quantum dot’s application in cell labeling.

4. Drug DiscoveryPatient Care

5. Biomarkers What Are Biomarkers? Concentration/ abundance Glucose electrolyte Cholesterol … Insulin PSA (prostate specific antigen) HER-2 (Breast cancer biomarker) CA-125 (Ovarian cancer biomarker) … ??? mg/mL ng/mL pg/mL

6. At this stage, cancel cells have to detected through ultra-sensitive biomarkers

13. Gold Nanoparticle and Its Application in Ultra-sensitive DNA and Protein Detection

14. TEM image of the cluster of DNA- functionalized gold nanoparticles Assembly of DNA-functionalized Gold Nanoparticles

15. Use functionalized Gold Nanoparticles to Detect DNA Color change of the gold colloidal solution reflect change of the cluster size Au…ATTCCGA TTAGGCT…Au …TAAGGCT…AATCCGA… DNA probe Sequences are designed so that the target DNA will link the particles together

16. Ultra-sensitive Detection of DNA with Nanoparticle assembly Array electrodestarget DNA Silver Staining The gold nanoparticles are linked together by target DNA Measure the resistance between 2 electrodes to tell the concentration of target DNA Sensitivity  500 fmol or 10 9 molecules.

17. Application of Ultra-sensitive DNA Detection Virus detection – HIV, SARS, Bird flu, etc. Biomarker detection – DNA/RNA related to cancer and drug metabolism. Personalized Medicine – Predict each patient’s response to certain treatment based on his/her gene.

18. Quantum Dot and Its Application in Cell labeling

19. What are Quantum Dots (QDot)? 655 605 585 565 525 nm Size of the nanocrystal determines the color Size is tunable from ~5-15 nm (±3%) Size distribution determines the spectral width Highly fluorescent, nanometer-size, single crystals of semiconductor materials

20. Size of Cells & Qdots

21. Qdot Conjugates are Engineered Core Nanocrystal (CdSe ) - Determines color Inorganic Shell (ZnS) – Improves brightness and stability Organic Coating - Provides water solubility and functional groups for conjugation. Biomolecules -Covalently attached to polymer shell - Immuoglobulins (Abs) - Streptavidin, Protein A - Receptor ligands - Oligonucleotides 15 - 18 nm

22. High level Her 2/neu expression in SK-BR-3 cells Quantum dots is up to 50x brighter. Low level of Her 2/neu expression in MDA-MB- 231 cells Organic dye is undetectable. Excellent Brightness and Photo-stability

23. Sharp and distinguishable peaks enable multi-color detection Minimal (<5%) cross-talk using 20nm bandpass filters

24. Example Application - Qtracker® Cell Labeling Kits Goal of Qtracker: To provide customers an approach to track live cells using Qdot-sav. cell + Qdot Why do we want to Tracking living cells? To understand cell differentiation To study cell-cell interaction To monitor cancer cells’ response to drug/ radiation treatment.

25. Qdot Delivered into Live Cells No Biotin10x Biotin 50X Biotin As the concentration of quantum dots increases, the signal strength from the cell increases

26. Cellular Retention Time of QDots The cells are still alive after multiple days. When the cells divide, the dots in the cell are divided also, causing signal to drop according to division rate.

27. Three-Color QDots Cell Labeling Different Qdots can be functionalized so that each dot can enter a specific type of cells. Enables multi-color labeling and tracking. Cell-cell interaction can be studied with the same approach.

28. “Scratch Assay” with QDots Cell Labeling Kit Cell’s migration speed can be measured by scratch an “X” mark in a cell culture, then monitor the time it takes for the cells to cross the boundary.

29. Other Qdot Applications Proteomics – Protein detection Research Cell Biology – cell tracking Pathology – tissue analysis Pharmaceutical R&D Products –High Content Screening –Mid Content Screening – Cell based assays –Multiplex Cellular Assays In vivo Imaging Agents