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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.

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nanomaterial s application in biotechnology

Nanomaterial’s Application in Biotechnology

Lei Sun

Project manager

Digital Health Group

Intel Corp.

outline
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.
slide4

Patient Care

Drug Discovery

what are 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

Biomarkers

ng/mL

pg/mL

slide14

Assembly of DNA-functionalized Gold Nanoparticles

TEM image of the cluster of DNA-functionalized gold nanoparticles

slide15

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

slide16

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
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
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

qdot conjugates are engineered

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
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
Sharp and distinguishable peaks enable multi-color detection

Minimal (<5%) cross-talk using 20nm bandpass filters

example application qtracker cell labeling kits

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
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
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
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
“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
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