Nanomaterial’s Application in Biotechnology Lei Sun Project manager Digital Health Group Intel Corp.
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.
Patient Care Drug Discovery
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 Biomarkers ng/mL pg/mL
At this stage, cancel cells have to detected through ultra-sensitive biomarkers
Gold Nanoparticle and Its Application in Ultra-sensitive DNA and Protein Detection
Assembly of DNA-functionalized Gold Nanoparticles TEM image of the cluster of DNA-functionalized gold nanoparticles
Use functionalized Gold Nanoparticles to Detect DNA Au…ATTCCGA TTAGGCT…Au …TAAGGCT…AATCCGA… DNA probe Sequences are designed so that the target DNA will link the particles together Color change of the gold colloidal solution reflect change of the cluster size
Ultra-sensitive Detection of DNA with Nanoparticle assembly Array The gold nanoparticles are linked together by target DNA electrodes target DNA Silver Staining Measure the resistance between 2 electrodes to tell the concentration of target DNA Sensitivity 500 fmol or 109 molecules.
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.
What are Quantum Dots (QDot)? Highly fluorescent, nanometer-size, single crystals of semiconductor materials 655605585565 525 nm Size of the nanocrystal determines the color Size is tunable from ~5-15 nm (±3%) Size distribution determines the spectral width
15 - 18 nm • Biomolecules • Covalently attached to polymer shell • Immuoglobulins (Abs) • Streptavidin, Protein A • Receptor ligands • Oligonucleotides 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.
Excellent Brightness and Photo-stability • 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.
Sharp and distinguishable peaks enable multi-color detection Minimal (<5%) cross-talk using 20nm bandpass filters
cell Qdot Example Application - Qtracker® Cell Labeling Kits Goal of Qtracker: To provide customers an approach to track live cells using Qdot-sav. + • 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.
Qdot Delivered into Live Cells No Biotin 10x Biotin 50X Biotin As the concentration of quantum dots increases, the signal strength from the cell increases
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.
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.
“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.
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