Nanomaterial’s Application in Biotechnology Lei Sun Project manager Digital Health Group Intel...
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Transcript of Nanomaterial’s Application in Biotechnology Lei Sun Project manager Digital Health Group Intel...
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.
Biomarkers
What Are Biomarkers? Concentration/
abundanceGlucoseelectrolyteCholesterol
…
Insulin
PSA (prostate specific antigen)
HER-2 (Breast cancer biomarker)
CA-125 (Ovarian cancer biomarker)
…
???
mg/mL
ng/mL
pg/mL
TEM image of the cluster of
DNA-functionalized gold nanoparticles
Assembly of DNA-functionalized Gold Nanoparticles
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
Ultra-sensitive Detection of DNA with Nanoparticle assembly Array
electrodes targetDNA
Silver Staining
The gold nanoparticles are linked together by
target DNA
Measure the resistance between 2 electrodes to tell the concentration of target DNASensitivity 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 What are Quantum Dots (QDot)?(QDot)?
655 605 585 565
525 nmSize 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
Qdot Conjugates are Qdot Conjugates are Engineered 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
•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-Excellent Brightness and Photo-stability stability
Sharp and distinguishable Sharp and distinguishable peaks enable multi-color peaks enable multi-color
detectiondetection
Minimal (<5%) cross-talk using 20nm bandpass
filters
Example Application - Example Application - Qtracker® Cell Labeling KitsQtracker® 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.
Qdot Delivered into Live CellsQdot 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 Cellular Retention Time of QDotsQDots
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 Three-Color QDots Cell LabelingLabeling
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 Scratch Assay” with QDots Cell Labeling KitCell 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.