나노생물학 chapter 1

Chapter1. Basics and Classification

of Nanobiotechnology
I. Nanomaterials

I-1. Nanoparticles
 General Properties of Nanoparticles
• Nanoparticles (NPs) form the bulk of nanomaterials.
• They can be made of different materials (e.g., metallic, semiconducting,
magnetic, organic, inorganic, and etc.)
• At those sizes, every particle has new properties that change depending
on its size, shape and compositions.
• As matter is shrunk to nanoscale, electronic, optical, and magnetic
properties change radically.
• Some applications of nanoparticles take advantage of the fact that more
surface area is exposed when material is broken down to smaller sizes.
 Nanoparticles
1. Metallic nanoparticles (gold, silver, etc…)
2. Cubosomes
3. Fullerenes
4. Lipoparticles
5. Nanoshells
6. Magnetic Nanoparticles
7. Polymer nanoparticles
8. Quantum dots
9. Silica nanoparticles
10. Nanoparticles assembly into micelles
1. Metallic nanoparticles (eg., Au, Ag)
• Mass spectrometry analysis has determined the formula of gold
nanocrystal molecules to be Au333(SR)79 (Qian et al. 2012 ).
• This metallic nanocrystal molecule exhibits fcc-crystallinity and surface
plasmon resonance (SPR) at approximately 520 nm.
• Simulations have revealed that atomic shell closing largely contributes to
the particular robustness of Au333(SR)79, albeit the number of free
electrons is also consistent with electron shell closing based on
calculations using a confined free electron model.
• Biocompatible
• Photostable
• Excellent optical property (absorption, scattering,...)
• Photothermal
• Easy synthesis and easy modification
• High spatial resolution
Metallic nanoparticles (eg., Au, Ag)

S. J. Tan et al., Nature Nanotechnology (2011)

 What if the size of metallic particle
becomes smaller than 100 nm ?
 Noble-metal nanoparticles with sizes smaller than the wavelength of visible light show strong
resonances for light scattering and absorption, due to the excitation of localized surface plasmons.
 At resonance, light resonantly drives collective oscillation of the conduction electrons of the metal
nanoparticle, which therefore acts as a radiating dipole.
 Its resonance frequency is strongly dependent on particle shape and dielectric environment,
which enables tuning of its "color” throughout the visible and into the near-infrared(IR) regime of
the spectrum, while keeping particle size well below 100nm.

 The ancient application: These effects are illustrated by the

historic stained glass which adorn medieval cathedrals, with
incorporating metal nanoparticle dopants.
 More modern applications, increasingly at a single-
nanoparticle level, are in (i) imaging probe by tagging of
biomolecules, (ii) enhancement of light emission from
nanoscale photon sources, and (iii) molecular sensing.

I. Surface Plasmon Resonance

II. Photothermal conversion
III. Surface Enhanced Effect
(1) Surface Plasmon Resonance (SPR)
(2) Photothermal Conversion
 Example:
Photothermal Therapy & Optogenetics
(3) Surface-enhanced Effect
 Examples:
SERS Active Arrays of Nanoantennae
 Examples:
Bioinspired Nanoparticles: gold plant viruses

Luke P. Lee’ s group

Light: Science & Applications 4, e267 (2015)
 2. Cubosomes
Nanomedicine, 5(9):1385-1399 (2005) Journal of Nanoparticle Research 4: 297-311 Biol. Pharm. Bull. 30(2) 350-353 (2007
(2002)

• Methods and compositions for producing lipid-based cubic phase nanoparticles were first discovered in the
1990s.
• Since then, a number of studies have described properties such as particle size, morphology, and stability of
cubic phase dispersions, which can be tuned by composition and processing conditions.
• Because of their unique microstructure, they are biologically compatible and capable of controlled release of
solubilized active ingredients such as drugs and proteins.
• As a drug delivery vehicle, high drug payloads, stabilization of peptides or proteins, and simple preparation
process are also advantages of a cubosome.
• The ability of cubic phase to incorporate and control release of drugs of varying size and polar characteristics,
and biodegradability of lipids make it a versatile drug delivery system for various routes of administration,
including oral, topical (or mucosal), and intravenous.
• Furthermore, proteins in cubic phase appear to retain their native conformation and bioactivity and are
protected against chemical and physical inactivation.
 3. Fullerenes
Giant fullerenes

• Fullerene technology derives from the discovery in 1985 of carbon-60, a molecule of 60 carbon
atoms that form a hollow sphere one nanometer in diameter.
• The molecule was named buckyball or fullerene or buckminsterfullerene, because of its
similarity to the geodesic dome designed by Buckminster Fuller.
• Subsequent studies have shown that fullerenes actually represent a family of related structures
containing 20, 40, 60, 70, or 84 carbons. C60, however, is the most abundant member of this
family.
• Fullerenes are entirely insoluble in water, but suitable functionalization makes the molecules
soluble. Initial studies on water-soluble fullerene derivatives led to the discovery of the
interaction of organic fullerenes with DNA, proteins, and living cells.
Fullerenes and Carbon Nanomaterials

Science 329(5996), 1159-1160, 2010

 4. Lipoparticles

http://www.integralmolecular.com

• Lipoparticles are nanometer-sized spheres surrounded by a lipid bilayer and embedded with
conformationally intact integral membrane proteins.
• Interactions with integral membrane proteins have been particularly difficult to study because the
proteins cannot be removed from the lipid membrane of a cell without disrupting the structure and
function of the protein.
• Lipoparticles provide an alternative to living cells, membrane preparations, and detergent-
solubilized proteins by offering concentrated membrane proteins in their native conformation.
• Lipoparticles are compatible with a wide variety of detection platforms and are ideal for drug and
antibody discovery and characterization.
– Immunization for antibody elicitation/ Phage and yeast display/ Antibody screening by ELISA/ Kinetic analysis
of antibody binding by biosensor/ Radioligand and fluorescent binding assays
 5. Nanoshells

• Nanoshells are ball-shaped structures measuring ~100 nm and consist of a core of nonconducting
glass that is covered by a metallic shell, which is typically gold or silver.
• The core/shell ratio and overall size of a gold nanoshell influence its scattering and absorption.
• By varying the relative dimensions of core and shell, the optical resonance of these nanoparticles
can be systematically varied over a broad region ranging from the near-UV to the mid-infrared.
• This range includes the NIR wavelength region where tissue transmissibility peaks, which forms the
basis of absorbing nanoshells in NIR thermal therapy of tumors.
• In addition to spectral tunability, nanoshells offer other advantages over conventional organic dyes
including improved optical properties and reduced susceptibility to chemical/thermal denaturation.
• Furthermore, the same conjugation protocols used to bind biomolecules to gold colloid are easily
modified for nanoshells.
• Nanoshells possess highly favorable optical and chemical properties for biomedical imaging and
therapeutic applications. These particles are also effective substrates for surface-enhanced Raman
scattering (SERS) and are easily conjugated to antibodies and other biomolecules.
• Several potential biomedical applications of nanoshells are under development, including
immunoassays, modulated drug delivery, photothermal cancer therapy, and imaging contrast
agents.
 Properties of Nanoshells
Naomi Halas’s Group. (Rice Univ.)

Transmission electron microscopy (TEM) images of nanoshell growth

phases from silica core (left) to gold-covered nanoshell (right)
Optical resonances of gold shell-silica core
nanoshells as a function of their core/shell ratio.

Technology in Cancer Research & Treatment, 3(1),33-40 (2004)

 6. Magnetic Nanoparticles
• Paramagnetic particles are important tools for cell sorting, protein separation, and single-
molecule measurements. The particles used in these applications must meet the following
requirements: uniform in size, highly paramagnetic, stable in physiological salt buffer,
functionizable.
• They have been used for the detection of model pathogens. Paramagnetic nanoparticles,
which are linked to antibodies, enable highly specific biological cell separations.
• Superparamagnetic iron oxide nanoparticles (SPION) with appropriate surface chemistry have
been widely used experimentally for numerous in vivo applications such as magnetic
resonance imaging (MRI) contrast enhancement, tissue repair, immunoassay, detoxification
of biological fluids, hyperthermia, drug delivery, in cell separation, etc.
• Magnetic nanoparticles can bind to drugs, proteins, enzymes, antibodies, or nucleotides and
can be directed to an organ, tissue, or tumor using an external magnetic field or can be
heated in alternating magnetic fields for use in hyperthermia.

출처: http://blog.naver.com/kims_pr/119589930 20
 Properties and Applications of
Magnetic Nanoparticles
• Paramagnetic complexes, which are usually gadolinium
(Gd3+) or manganese (Mn2+) chelates, accelerate
longitudinal (T1) relaxation of water protons and exert
bright contrast in regions where the complexes localize.
 Gadolinium diethylenetriaminepentaacetate (Gd-
DTPA) has been the most widely used of such
complexes and its main clinical applications are
focused on detecting the breakage of the blood-
brain barrier (BBB) and changes in vascularity,
flow dynamics, and perfusion.
 Manganese-enhanced MRI (MEMRI), which uses
manganese ion (Mn2+) as a T1 contrast agent, is
applicable to animals only owing to the toxicity of
Mn2+ when it accumulates excessively in tissues
and despite the increasing appreciation of this
technique in neuroscience research.

Chem. Commun., 1203-1214 (2007)

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 Properties and Applications of
Magnetic Nanoparticles
Imaging of brain structures by using MnO nanoparticles

Figure 1. a) TEM images of water-dispersible MnO

nanoparticles with particle sizes of 7, 15, 20, and 25 nm. b) T1-
weighted MR image of MnO nanoparticles from a 3.0 T clinical
MRI system.
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Angew. Chem. 119, 5493 –5497 (2007)
 7. Polymer Nanoparticles
• Polymer nanoparticles, synthetic as well as biopolymers, are biocompatible, biodegradable,
and nontoxic.
• They can be conjugated with other nanoparticles. Different types of polymer nanoparticles
have been designed as drug delivery devices.
• Biodegradable polymeric nanoparticles are promising drug delivery devices because of their
ability to deliver proteins, peptides, and genes as well as targeting therapeutics to specific
organs/tissues.
• Although several synthetic polymers are available, natural polymers are still popular for drug
delivery; these include acacia gum, chitosan, gelatin, and albumin.
• Examples of synthetic biodegradable polymers for controlled release drug delivery are
polylactides (PLA), polyglycolides(PLG), and poly(lactide-co-glycolides) or PLGA.

• Poly(lactic-co-glycolic acid) (PLGA) is one of

the most successfully used biodegradable
polymers because its hydrolysis leads to
metabolite monomers, lactic acid and
glycolic acid.

Hydrolysis of PLGA 23
 Application of Polymer Nanoparticles
Development of Dtxl-encapsulated pegylated PLGA NP-Apt bioconjugates.

Farokhzad O C et al. PNAS 2006;103:6315-6320 24

 8. Quantum Dots
• Quantum dots (QDs) are nanoscale crystals of semiconductor material that glow or fluoresce
when excited by a light source such as a laser.
• They are composed of 200–10,000 atoms. These semiconductor materials can be made from an
element, such as silicon or germanium, or a compound, such as cadmium selenide (CdSe) or zinc
sulfide (ZnS) or cadmium sulfide (CdS).
• The size of the QD determines the frequency of light emitted when irradiated with low energy
light (size-tunable emission and simultaneous excitation).  Multicolor optical coding for
biological assays
• Current dyes used for lighting up protein and DNA fade quickly, but QDs could allow tracking of
biological reactions in living cells for days or longer.
• Limitations: The QDs were initially found to be unstable and toxic and difficult to use in solution.
UV illumination

Ambient illumination

Analyst 129(8): 672-7 (2004)

http://education.mrsec.wisc.edu/background/quantum_dots/  Size-tunable fluorescence spectra of CdSe quantum dots (A), and
 Collection of CdSe quantum dot nanoparticles of different size illustration of the relative particle sizes (B). From left to right, the
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: Solutions are in order of increasing particle size (longer growth time). particle diameters are 2.1 nm, 2.5 nm, 2.9 nm, 4.7 nm, and 7.5 nm.
 The core/shell CdSe/CdS quantum dot imaged on this picture has a
diameter of 12nm. It has been taken with a Transmission Electron
Microscope at magnification 400,000x. Due to a favorable orientation of
the crystal axes, one can distinguish the electron-opaque atoms.

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(https://blog.espci.fr/qdots/2011/01/12/a-coreshell-cdsecds-quantum-dot/)
 Applications of QDs
• Due to their sheer brightness and high photostability, QDs have the ability
to act as molecular beacons. When attached to compounds or proteins of
interest, QDs enable researchers to track movements within biological
media or whole organisms, significantly impacting the way medical
professionals study, diagnose, and treat diseases.

• Applications of QDs include the following:

– Life sciences research: tracking proteins in living cells
– Fluorescence detection: microscopy, biosensors, multicolor flow cytometry
– Molecular diagnostics
– Ex vivo live cell imaging
– In vivo targeting of cells, tissues, and tumors with monitoring by PET and MRI
– High-throughput screening
– Identification of lymph nodes in live animals by NIR emission during surgery

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 9. Silica Nanoparticles
• In the case of silica, the formation of diatom shell or sponge spicule has attracted
much attention in the last decade since it could provide key information to
elaborate new hierarchically structured materials and nanodevices.
• The mineral phase is thought to be formed by the controlled assembly of
nanoparticles generated in vivo from diluted precursor solutions, in the presence
of biomolecular templates.
• Biomimetic approaches have led to the identification of several natural or
synthetic molecules that are able to activate silica formation in conditions that
closely resemble those found in the living organisms’ intracellular compartments.
• Extraction and characterization of biosilicifying molecules from living organisms,
however, are still limited.
• Currently, silica nanopaticles can be synthesize by sol-gel methods without
biotemplates.
• Advantages: biodegradable.
• Applications: drug delivery and gene therapy.
Biosilicification

- Extraction: silica particles formed from

sodium silicate solutions in the presence
of bovine serum albumin (left) and
gelatine (right).

- Templating: SEM images of silica coating

on alginate/p-Lys (left panel) and on the
surface of gelatine capsule (right panel).

Current Nanoscience, 2005, 1, 00-00 1

“Mimicking Biogenic Silica Nanostructures Formation”
 10. Nanoparticles Assembly into
Micelles
- Assembly of gold and silver nanoparticle
building blocks into larger structures is based on
a novel method that goes back to one of
nature’s oldest known chemical innovations, i.e.,
the self-assembly of lipid membranes that
surround every living cell.
- The method makes use of the hydrophobic
effect, a biochemical phenomenon that all living
creatures use to create membranes, ultrathin
barriers of fatty acids that form a strong, yet
dynamic, sack around the cell, sealing it from
the outside world.
- Cell membranes are one example of a micelle, a
strong bilayer covering that is made of two
sheets of lipid-based amphiphiles molecules.
- All micelles form in three shapes: spheres,
cylinders, and sack-like vesicles.
J. AM. CHEM. SOC. 2006, 128, 15098-15099
Shapes of Micelles

-Schematic representation of organized aggregates of

surfactants: (a) normal micelles, (b) reverse micelles,
(c) cylindrical micelle, (d) planar lamellar phases, (e)
onion-like lamellar phases and (f) interconnected
cylinders.
Chem. Soc. Rev., 2010, 39, 474-485