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Nanotechnology Creation of useful materials, devices, and systems through the manipulation of matter on nanometer scale. - Generally nanotechnology deals with structures sized between 1 to 100 nanometer in at least one dimension. Ability to design systems with defined structure and function on the nanometer scale. Involves developing materials, devices within that size, and analytical tools (methodology), which can be used for analysis and measurement on a molecular scale Interdisciplinary area : Biology, Physics, Chemistry, Material science, Electronics, Chemical Engineering, Information technology Nanotechnology Plays by Different Rules Normal scale Nanoscale Analytical methods and Nano-sized materials Analytical tools : Atomic force microscopy(AFM), Electron microscopy (EM) Nano-sized materials Unusual and different property - Semiconductor nanocrystals: Size-dependent optical property - Nanoparticles: Magnetic nanoparticles (Ferromagnetic, superparamagnetic), Gold, Carbon nanotubes, Quantum dots (Semiconductor nanocrystals) Future implications of nanotech Nanotechnology may be able to create many new materials and devices with a vast range of applications, such as in medicine, biomaterials, electronics, and energy production. Nanotechnology raises many of the same issues as with any introduction of new technology, including concerns about the toxicity and environmental impact of nanomaterials, and their potential effects on global economics. Nano-Biotechnology Integration of nano-sized/structured materials, nanoscale analytical tools, and nano-devices into biological sciences for development of new biomaterials and analytical toolkits as well as for understanding life science Use of bio-inspired molecules or materials Typical characteristics of Biological events/materials - Self assembly - Highly efficient : high energy yield - Very specific : extremely precise Bio-molecules Proteins, DNA, RNA, Aptamers, Peptides, Antibody, Virus Nano-Bio Convergence Bio-inspired device and system Bio-Technology Nano-Technol Molecular Imaging Molecular Switch DNA barcode Biochip / Biosensor Nanotherapy / Delivery Bionano-machine / Nano-Robot Applications and Perspectives of Nanobiotechnology Development of tools and methods - More sensitive - More specific - Multiplexed - More efficient and economic Implementation Diagnosis and treatment of diseases - Rapid and sensitive detection (Biomarkers, Imaging) - Targeted delivery of therapeutics Drug development - Understanding of life science Examples Nano-Biodevices Nano-Biosensors Imaging with nanoparticles Analysis of a single molecule/ a single cell Issues to be considered Synthesis or selection of nano-sized/ structured materials Functionalization with biomolecules or for biocompatibility Integration with devices and/or analytical tools Assessment : Reproducibility, Toxicity Implementation The size of Things NanoBiotech was initiated by the development of AFM that enables imaging at atomic level in 1980 AFM (Atomic Force Microscope) One of the foremost tools for imaging, measuring, and manipulating matters at the nanoscale. A cantilever with a sharp tip (probe) at its end that is used to scan the specimen surface When the top is brought into a close proximity of a sample surface, force between the tip and sample leads to a deflection of the cantilever according to Hooke’s law Deflection is measured using a laser spot reflected from the top surface of the cantilever into an array of photodiode VEECO TESPA® VEECO TESPA-HAR® NANOWORLD SuperSharpSilicon® Tip length : 10 m Radius : 15~20 nm Tip length :10 m (last 2 m 7:1) Radius : 4~10 nm Tip length :10 m Radius : 2 nm Example showing the resolution of protein structure by AFM Image of ATP synthase composed of 14subunits Molecular imaging Biomedical & Biological Sciences : Ultra-sensitive imaging of biological targets under non-invasive in-vivo conditions Fluorescence, positron emission tomography, Magnetic resonance imaging Ultra-sensitive imaging - Cancer detection, cell migration, gene expression, localization of proteins, angiogenesis, apotosis - MRI : Powerful imaging tool as a result of non-invasive nature, high spatial resolution and tomographic capability Resolution is highly dependent on the molecular imaging agents Signal enhancement by using contrast agents : iron oxide nanoparticles Semiconductor Nanocrystals Quantum Dots - Properties and Biological Applications Synthesis of CdSe/ZnS (Core/Shell) QDs Step 1 CdO + Se CdSe Solvent : TOPO, HAD, TOP Surfactant : TDPA, dioctylamine Step 2 CdSe/ZnS 5.5 nm (red) ZnS ZnEt2 + S(TMS)2 CdSe Growth temperature 140 ℃ (green) 200 ℃ (red) Ar Thermocouple Se solution CdO solution 320 ℃ 20 nm Bawendi et al. J. Am. Chem. Soc. (1994) Optical Properties Of Quantum Dots a) Multiple colors b) Photostability c) Wide absorption and narrow emission d) High quantum yield Quantum Yield ≥ 60 ~ 70 % Single source excitation QD Surface Coating for Biocompatibility ① NH2 NH2 Encapsulation with the hydrophobic core of a micell NH2 + Coating with PC P P O O P O CdSe O P OP O O P P CdSe QDs N + P P O O ZnS P O OP ZnS O O P Coating of the outer shell with ZnS OP OP O P O P NH2 OP CdSe CdSe NH2 N + OP O P O N NH2 P + N P CdSe/ZnS core-shell + N NH2 NH2 Quantum Dots Encapsulated in Phospholipid Micelles PEG-PE (n-poly(ethylenglycol)phosphatidylethanolamine): micell-forming hydrophilic polymer-grafted lipids comparable to natural lipoproteins PEG : low immunogenic and antigenic, low non-specific protein binding PC : Phosphatidylcholine Dubertret et al. Science (2002) In Vivo Cell Imaging Y + QD Organelle QD-Antibody conjugates Antigen Y ▲ 3T3 cell nucleus stained with red QDs and microtubules with green QDs QD Organelle - Multiple Color Imaging - Stronger Signals Wu et al. Nature Biotech. 2003 21 41 In Vivo Cell Imaging Live Cell Imaging Quantum Dot Injection ▶ Red Quantum Dot locating a tumor in a live mouse Cell Motility Imaging 10um ◀ Green QD filled vesicles move toward to nucleus (yellow arrow) in breast tumor cell Alivisatos et al., Adv. Mater., 2002 14 882 Biosystems • Inherent capabilities of molecular recognition and self-assembly • Attractive template for constructing and organizing the nano-structures • Proteins, toxin, coat proteins of virus etc. α -Hemolysin: Self-Assembling Transmembrane Pore A self-assembling bacterial exo-toxin produced by some pathogens like Staphylococcus aureus as a way to obtain nutrients lysis of red blood cells Alpha-hemolysin monomers bind to the outer membrane of susceptible cells. Monomers oligomerize to form a water-filled heptameric transmembrane channel that facilitates uncontrolled permeation of water, ions, and small organic molecules. Rapid discharge of vital molecules, such as ATP, dissipation of the membrane potential and ionic gradients, and irreversible osmotic swelling leading to the cell wall rupture (lysis), can cause death of the host cell. - Mushroom-like shape with a 50 A beta-barrel stem - Narrowest part (1.4 nm in diameter) of channel at the base of stem Biotechnological applications :Stochastic sensors A molecular adaptor is placed inside its engineered stem, influencing the transmembrane ionic current induced by an applied voltage Reversible binding of analytes to the molecular adaptor transiently reduces the ionic current - Magnitude of the current reduction : type of analyte - Frequency of current reduction intervals: analyte concentration Stochastic Sensors a : Histidine captured metal ions (Zn+2, Co+2, mixture ) b: CD captures anions (promethazine, imipramine, mixture) c : biotin ligand DNA sequencing Transmembrane pore can conduct big (tens of kDa) linear macromolecules like DNA or RNA Eelectrophoretically-driven translocation of a 58-nucleotide DNA strand through the transmembrane pore of alpha-hemolysin Changes in the ionic current by the chemical structure of individual strands Nucleotide sequence directly from a DNA or RNA strand A single nucleotide resolution Understanding Cancer and Related Topics Understanding Nanodevices Developed by: Jennifer Michalowski, M.S. Donna Kerrigan, M.S. Jeanne Kelly Brian Hollen Explains nanotechnology and its potential to improve cancer detection, diagnosis, and treatment. Illustrates several nanotechnology tools in development, including nanopores, quantum dots, and dendrimers. These PowerPoint slides are not locked files. You can mix and match slides from different tutorials as you prepare your own lectures. In the Notes section, you will find explanations of the graphics. The art in this tutorial is copyrighted and may not be reused for commercial gain. Please do not remove the NCI logo or the copyright mark from any slide. These tutorials may be copied only if they are distributed free of charge for educational purposes. What Is NanoBiotechnology? Water molecule Nanodevices Nanopores Dendrimers Nanotubes Quantum dots Nanoshells White blood cell A period Tennis ball Designing Nanodevices for Use in the Body Too Small Too Big Manufacturing Nanodevices X-ray beam Crystal Crystal Scattered X-rays Detector Nanodevices Atoms in crystal White blood cell Nanodevices Are Small Enough to Enter Cells Cell Nanodevices Nanodevices Water molecule White blood cell Nanodevices Can Improve Cancer Detection and Diagnosis NanoBiotechnology Imaging Physical Exam, Symptoms Nanodevices Can Improve Sensitivity Nanodevices could potentially enter cells Precancerous cells Normal cells and determine which cells are cancerous or precancerous. Precancerous cells Normal cells Nanodevices Can Preserve Patients’ Samples Traditional Tests Cells from patient Cells altered Active state lost Nanotechnology Tests Cells from patient Cells preserved Active state preserved Additional tests Nanodevices Can Make Cancer Tests Faster and More Efficient Patient A Patient B Cantilevers Can Make Cancer Tests Faster and More Efficient Cancer cell Antibodies with proteins Antibodies Bent cantilever Water molecule White blood cell Nanodevices Cantilevers Nanopores Single-stranded DNA molecule A T C G Nanopore Singlestranded DNA molecule A Nanopore T Nanopore Single-stranded DNA molecule Water molecule Nanodevices Nanopores White blood cell Quantum Dots Ultraviolet light off Quantum dot bead Water molecule Ultraviolet light on Quantum dots Nanodevices Quantum dots Quantum dots emit light White blood cell Quantum Dots Can Find Cancer Signatures Quantum dot beads Cancer cells Healthy cells Cancer cells Quantum dot beads Healthy cells Improving Cancer Treatment Traditional Treatment Drugs Nanotechnology Treatment Toxins Nanodevices Cancer cells Cancer cells Noncancerous cells Dead cancer cells Dead noncancerous cells Toxins Noncancerous cells Dead cancer cells Intact noncancerous cells Nanoshells Near-infrared light off Nanoshell Near-infrared light on Gold Nanoshell absorbs heat Water molecule Nanodevices Nanoshells White blood cell Nanoshells as Cancer Therapy Nanoshells Nanoshells Cancer cells Cancer cells Healthy cells Healthy cells Near-infrared light Dead cancer cells Intact healthy cells Nanodevices as a Link Between Detection, Diagnosis, and Treatment Traditional Cancer Treatment NanoBiotechnology Cancer Treatment Cancer cell Cancer cell Nanodevice Drug Imaging Reporting Detection Targeting Dendrimers Cancer cell Dendrimer Water molecule Nanodevices Dendrimer White blood cell Dendrimers : Highly Branched Dendritic Macromolecules Poly (amido amine) Dendrimers ● Characteristics Monodisperse macromolecule Globular (Spherical) Facile surface bio-functionalization Similar molecular size to biomolecules (Glucose oxidase 65.27.7 nm) ● Applications Vehicles for delivery of genes and drugs 4.5 nm G4 Poly(amidoamine) Dendrimer Biomimetic catalysts (Peptides-, Glycodendrimers) Medical applications (MRI contrast enhancer) Molecular carriers for chemical catalysts (Core, Peripheral) Dendrimers as Cancer Therapy Manipulate dendrimers to release their contents only in the presence of certain trigger (molecules or light) caged molecules Therapeutic agent Cancer detector Cell death monitor Reporter Water molecule Nanodevices Dendrimer White blood cell NanoBiotechnologies in Patient Care Today 2020 We would like to hear from you . . . 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