Lecture note

1. NANOBIOTECHNOLOGY
Mobt 4075
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2. Nanotechnology
• Atomic and molecular level study
• Structures sized between 1 to 100 nanometer in
at least one dimension
• Developing or modifying materials or devices
within that size
• Novel properties
• Components should remain at nanometer scale
• Involves imaging, measuring, modeling, and
manipulating matter at this length scale
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3. 1. Introduction
 Nanotechnology: It involves research and technology
development at the atomic, molecular or macro-molecular level in
the length scale of approximately 1 to 100 nm range.
 Biotechnology: Biotechnology is the use of biological processes,
organisms, or systems to manufacture products intended to
improve the quality of human life.
 The interface of these two worlds lies Nanobiotechnology
– It uses nanotechnology to analyse and create biological nanosystems
– It uses biological materials and structural plans to produce technical,
functional nanosystems.
• Nanobiotechnology is an emerging field at the crossroads of biotechnology and
material science and involved in many disciplines including physists, chemists,
engineers, information technologists, and material scientists as well as
biologists.
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6. History of Nanoparticles
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8. Major developments in the application of Nanotechnology
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12. Nanoparticles
• Nanoparticles are defined as particles that have at least one dimension in
the nanorange (1 to 100nm).
• Nanoscale materials (nanoparticles, nanopores, nanoshells, nanostructures
etc) allow highly sensitive detection by specific interactions with various
biomolecules on both surface and inside the cells.
• Nanotechnology helps in development of small, highly-efficient and
inexpensive sensors, with broad applications.
• These offer significant advantages over conventional sensors. This includes
greater sensitivity and selectivity, lower production costs, reduced power
consumption as well as improved stability.
• Because of their submicron dimensions, nanosensors, nanoprobes & other
nanosystems have allowed simple & rapid analyses in vivo.
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13. Nanomaterials….
• Biological systems often feature natural,
functional nanomaterials.
• The structure of foraminifera, viruses
(capsid), the wax crystals covering a
lotus or nasturtium leaf, spider-mite
silk are few examples of natural
nanomaterials.
• Natural inorganic nanomaterials occur
through crystal growth in the diverse
chemical conditions of the earth‘s crust.
Forex. clays display complex
nanostructures due to anisotropy of
their underlying crystal structure, &
volcanic activity can give rise to opals,
which are an instance of a naturally
occurring photonic crystals due to their
nanoscale structure.
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15. Nanoparticles
 Nanoparticles are the particles of size between 1nm to 100nm range).
 Nanometer - One billionth (10-9) of a meter
The size of Hydrogen atom 0.04 nm
The size of Proteins ~ 1-20 nm
Feature size of computer chips 180 nm
Diameter of human hair ~ 10 µm
 At the nanoscale, the physical, chemical, and biological
properties of materials differ in fundamental and valuable
ways from the properties of individual atoms and molecules or
bulk matter
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17. Why NANO..?
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19. Novel Properties of nanoparticles
• Small size
• High surface area
• Ease to suspend in liquids
• Deep access to cells and organelles
• Improved physical, chemical & biological
properties
Properties of nanoparticles are different from their
bulk counterparts.
Extremely high surface area to volume ratio results
in surface dependent material properties.
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20. Nano-scale effects on properties over conventional methods
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22. Nanomaterials
• Nanomaterials are commonly defined as materials with an average
grain size less than 100 nm.
• Nano-biomaterials display distinct biological effects when compared
with bulk materials having same chemical composition.
• Nanomaterials with fast ion transport are related also to nanoionics &
nanoelectronics
• Their nanoscaled size emanates novel characteristics such
as increased strength, chemical reactivity or conductivity.
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• Engineered nanomaterials (ENM) are materials created by
manipulation of matter at the nanoscale to produce new
materials, structures, and devices.
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23. Classification of Nanomaterials
Nanomaterials are classified according to the length scale of each of its
dimension:
• 0 D :zero scale all three dimensions in the nanoscale (nanoparticles).
• 1 D : one dimension in nanoscale and other two in macroscale ( nanofibers,
nanowires)
• 2 D : two dimensions in nanoscale and the other in the macroscale ( nano
sheets, thin films)
• 3 D : no dimensions at the nanoscale, all are in the macroscale
(nanostructures with nanomaterials
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25. Nanotools and Nanodevices
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32. Quantum Confinement
Quantum Confinement is the spatial confinement of
electron-hole pairs (excitons) in one or more
dimensions within a material.
o 1D confinement: Quantum Wells
o 2D confinement: Quantum Wire
o 3D confinement: Quantum Dot
• Quantum confinement is more prominent in
semiconductors because they have an energy gap in
their electronic band structure.
• Metals do not have a bandgap, so quantum size effects
are less prevalent. Quantum confinement is only
observed at dimensions below 2 nm.
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Therefore, the more spatially confined and localized a particle becomes, the
broader the range of its momentum/energy.
• This is manifested as an increase in the average energy of electrons in the
conduction band = increased energy level spacing = larger band gap

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39. Applications of QDs
• Quantum dots are
tiny crystals tha
glow/ fluoresce
when they are
stimulated by
ultraviolet light.
• Fluorescent
nanocrystals.
• Common QDs: CdS,
• CdSe, PbS, PbSe,
PbTd, CuCl
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50. Light emitters
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New applications for QDs are continuously being discovered.
• For example: Solar cells that incorporate QDs may lead to more efficient light
harvesting and energy conversion.

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Drug delivery, & cancer treatment

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69. IMPROVING MRI Magnetic resonance imaging)
• Iron oxide nanoparticles can used to improve
Magnetic Resonance Imagining (MRI) images of
cancer tumors.
• The nanoparticle is coated with a peptide that
binds to a cancer tumor, once the nanoparticles
are attached to the tumor the magnetic property
of the iron oxide enhances the images from the
Magnetic Resonance Imagining scan.
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70. Characterization of
nanomaterials
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72. Characterization of Nanoparticles
1. Size and surface Morphology
2. Specific Surface Area
3. Surface Charge and Electrophoretic Mobility
4. Surface Hydrophobicity
5. Density
6. Molecular weight Measurements of Nanoparticles
7. Drug Entrapment efficiency
8. Kinetic Study
9. Stability of Nanoparticles
10. Drug-Excipient compatibility studies
11. In-vitro Release Studies
12. Lamellarity
13. Phase Behaviour
14. Chemical Characterization (Liposomes)
15. Biological Characterization (Liposomes)
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73. A. Dynamic Light Scattering (DLS)-
DLS measures brownian motion and
relates this to the size of the particles
(Hydrodynamic diameter).
Bias toward larger particles.
We can determine polydispersity
index (PDI), zeta potential and
aggregation of particles.
Instrumentation - Zetasizer (Malvern
panalytical tnstrument, UK), Laser
source, Photon detector, Polystyrene
cuvettes/Quartz or optical quality
glass cuvettes with caps.
Dispersant –Water or whatever the
dispersant used is.
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74. B. Nano Sight (NTA)-
• Nano sight helps in
visualization and measuring
nanoparticle size &
• concentration with
precision and accuracy.
• Nano sight instrument uses
Nanoparticle Tracking
Analysis (NTA) to
characterize nanoparticles
from 10 nm – 2000 nm in
solution.
• Characterization of
aggregation state.
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75. C. Scanning Electron
Microscopy (SEM)-
SEM is used to visualize
the surface morphology
of organisms, cells and
materials.
Resolution is 1-2 nm.
Can determine the
elemental composition.
Determine the size,
shape, surface
morphology.
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76. D. Transmission Electron
Microscopy (TEM)-
Resolution is 0.1 – 0.2 nm.
Determine the internal
structure or arrangements
of the particles.
Measure the size, size
distribution, and
morphology.
Samples are prepared for
imaging by drying
nanoparticles on a grid that
is coated with a thin layer of
carbon/formvar.
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81. APPLICATIONS OF X-RAY DIFFRACTION
• Obtain XRD patterns are used to measure d-
spacings of the given compound.
• XRD is used to determination of Cis-Trans
isomerism.
• X-ray diffraction is used to measure thickness of
thin films and multi-layers.
• XRD is used to determine atomic arrangement.
• XRD is used to measure the size, shape and
internal stress of small crystalline regions.
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97. Infrared waves
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100. Synthesis of Nanoparticles
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101. Synthesis of Nanoparticles
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Top Down approach
These seek to create smaller devices by using larger
ones to direct their assembly
The most common top-down approach to
fabrication involves lithographic patterning
techniques using short wavelength optical sources
Bottom up Approach
These seek to arrange smaller components into more
complex assemblies
Use chemical or physical forces operating at the
nanoscale to assemble basic units into larger structures
examples :
1. Indiun gallium arsenide(InGaAs) quantum dots can be
formed by growing thin layers of InGaAs on GaAs
2. Formation of carbon nanotubes

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3 methods of synthesis of NP
1. Physical
2. Chemical
3. Biological

106. 1. Physical methods
2 physical methods: mechanical and vapor
I. Mechanical
1. High energy ball milling
2. Melt mixing
II. Vapour
1. Physical vapour deposition
2. Laser ablation
3. Sputter deposition
4. Electric arc deposition
5. Ion implantation
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108. 2. CHEMICAL METHODS OF SYNTHESIS
• Simple techniques
• Inexpensive instrumentation
• Low temperature (<350ºC)
synthesis
• Doping of foreign atoms (ions)
is possible during
• synthesis
• Large quantities of material
can be obtained
• Variety of sizes and shapes are
possible
• Self assembly or patterning is
possible
• Sol-gel method
• Pyrolysis/thermolysis
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116. Pyrolysis
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118. sol-gel Method
• The sol-gel process is a wet-chemical technique (also
known as chemical solution deposition) widely used
recently in the fields of materials science and ceramic
engineering.
Steps
• Formation of stable sol.
• Gelation
• Gel aging into a solid mass. This causes contraction of
the gel network, also phase transformations and
Ostwald ripening.
• Drying of the gel to remove liquid phases. This can lead
to fundamental changes in the structure of the gel.
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119. Sol-gel Method…
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120. Advantages of sol-gel Method
• 2 types of materials or components- “sol” and
“gel”
• M. Ebelman synthesized them in 1845
• Low temperature process- less energy
consumption and less pollution
• Generates highly pure, well controlled ceramics
• Economical route, provided precursors are not
expensive
• Possible to synthesize nanoparticles, nanorods,
nanotubes etc.,
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121. COLLOIDS AND COLLOIDS IN SOLUTION
• Nanoparticles synthesized by chemical
methods form “colloids”
• Two or more phases (solid, liquid or gas) of
same or different materials co-exist with the
dimensions of at least one of the phases less
than a micrometre
• May be particles, plates or fibres
• Nanomaterials are a subclass of colloids, in
which the dimensions of colloids is in the
nanometre range
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123. 3. BIOLOGICAL/Green METHODS
• Green synthesis
3 types:
1.Use of microorganisms like fungi,
yeats(eukaryotes) or bacteria,
actinomycetes(prokaryotes)
2. Use of plant extracts or enzymes
3.Use of templates like DNA, membranes,
viruses and diatoms
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124. SYNTHESIS USING MICROORGANISMS
• Microorganisms are capable of interacting with metals coming in
contact with hem through their cells and form nanoparticles.
• The cell- metal interactions are quite complex
• Certain microorganisms are capable of separating metal ions.
• Pseudomonas stuzeri Ag259 bacteria are commonly found in silver
mines.
• Capable of accumulating silver inside or outside their cell
• walls
• Numerous types of silver nanoparticles of different shapes can be
produced having size <200nm intracellularly
• Low concentrations of metal ions (Au⁺,Ag⁺ etc) can be converted to
metal nanoparticles by Lactobacillus strain present in butter milk.
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125. • Fungi – Fusarium oxysporum challenged with gold or silver salt for
app. 3 days produces gold or silver nanoparticles extracellularly.
• Extremophilic actinomycete Thermomonospora sp. Produces gold
nanoparticles extracellularly.
• Semiconductor nanoparticles like CdS, ZnS, PbS etc., can be
produced using different microbial routes.
• Sulphate reducing bateria of the family Desulfobacteriaceae can
form 2-5nm ZnS nanoparticle. Klebsiella pneumoniae can be used to
synthesize CdS nanoparticles.
• when [Cd(NO₃)₂] salt is mixed in a solution containing bacteria and
solution is shaken for about1 day at ~38ºC ,CdS nanoparticle in the
size range ~5 to 200 nm can be formed.
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126. SYNTHESIS USING PLANT EXTRACTS
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129. SYNTHESIS USING DNA
• CdS or other sulfide nanoparticles can be synthesized using DNA.
• DNA can bind to the surface of growing nanoparticles.
• ds Salmon sperm DNA can be sheared to an average size of 500bp.
• Cadmium acetate is added to a desired medium like water, ethanol,
propanol etc.
• Reaction is carried out in a glass flask- facility to purge the solution
and flow with an inert gas like N₂.
• Addition of DNA should be made and then Na₂S can be added
dropwise.
• Depending on the concentrations of cadmium acetate, sodium
chloride and DNA ,nanoparticles of CdS with sizes less than ~10 nm
can be obtained.
• DNA bonds through its negatively charged PO₄ group to positively
charged (Cd+) nanoparticle surface.
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130. USE OF PROTEINS, TEMPLATES LIKE DNA , S- LAYERS ETC
• Various inorganic materials such as
carbonates, phosphates, silicates etc are
found in parts of bones, teeth, shells etc.
• Biological systems are capable of integrating
with inorganic materials
• Widely used to synthesize nanoparticles
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131. FERRITIN
• Ferritin is a colloidal protein of nanosize.
• Stored iron in metabolic process and is abundant
in animals.
• Capable of forming 3 dimensional hierarchical
structure.
• 24 peptide subunits – arranged in such a way that
they create a central cavity of ~6 nm.
• Diameter of polypeptide shell is 12 nm.
• Ferritin can accommodate 4500 Fe atoms.
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134. PROCEDURE TO CONVERT FERRITIN TO
APOFERRITIN
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135. Applications of Nanomaterials
and Nanoparticles
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136. Application of nanopaticles and nanomaterials
Application on many fields such as:
o Medicine/Health : Nanomedicine
o Food & agriculture
o Biotechnology
o Information technology
o Mechanical engineering & Robotics
o Advance materials & textiles
o Energy and Environment
o National security & defence
o Aerospace
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158. Multiplex Diagnosis
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 Four quantum dots of different diameter (i.e. different color) are respectively
functionalized with four different antigens. Allowing for the distinction of two
distinct phenotypes.

159. Cancer Therapy
 There is a search dual-mode nanoparticle that can detect a
tumor (imaging)and destroy it (therapy).
 There is two action modes for therapeutical nanoparticles.
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Passive Targeting Active Targeting
Based on nanoparticle
functionalization for specific
targeting of cancerous cells
Based on retention effect of
particle of certain hydrodynamic
size in cancerous tissues
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160. Taking advantage of retention
 Nanoparticles injected in the
blood stream do not permeate
through healthy tissues.
 Blood vessels in the surrounding
of tumorous tissues are defective
and porous.
 injected in the blood permeate
through blood vessels toward
tumorous tissues, wherein they
accumulate.
 Tumorous tissues suffer of
Enhanced Permeability and
Retention effect.
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161. Respirocyte- A proposed nanorobot
Respirocytes are:
 Artificial mechanical red blood
cells.
 Carry oxygen and carbon dioxide
molecules.
 Deliver 236 times more oxygen to
the body tissues when compared
to natural red blood cells .
Applications :
– Treatment of Anemia
– Transfusions and perfusions
– Fetal and Child Related
disorders
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• Spherical 1 micro meter diameter sized
• Constructed of 18 billion atoms

162. Lab-on-a-Chip
The Ideal Technology for Bio-chemical Analysis
• A lab-on-a-chip (LOC) is a device that integrates one or
several laboratory functions on a single chip of only
millimeters to a few square centimeters in size.
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163. What can “Lab-on-a-chip” do?
 Biochemical assays: real-time PCR, immunoassay,
dielectrophoresis for detecting cancer cells and bacteria, etc.
 Chemical application: separating molecules from mixtures,
chemical reactors, chemical detections etc.
 Biological application: cell coculture, biosensor, drug
screening, single-cell analysis, etc.
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164. Disadvantages of LOCs
 Novel technology and therefore not yet fully developed.
 Processes in LOCs more complex than in conventional lab
equipment.
 Detection principles may not always scale down in a positive
way, leading to low signal-to-noise ratios.
 Although the absolute geometric accuracies and precision in
microfabrication are high, they are often rather poor in a
relative way, compared to precision engineering for instance.
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165. Nanobiosensor (Biochip)
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166. Types of Nanosensors based on applications
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175. Nanotechnology in Agriculture
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176. Nanotech Delivery Systems for Pests, Nutrients,
& Plant Hormones
• Nanosensors dispersed in the field can also detect the
presence of plant viruses and the level of soil
nutrients.
• Nano encapsulated slow release fertilizers have also
become a trend to save fertilizer consumption, & to
minimize environmental pollution.
• Nanobarcodes and Nano processing could also be
used to monitor the quality of agricultural products.
• Used to study the effect on PGRs especially Auxin*.
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177. Nanoparticles and Recycling Agricultural Waste
• In cotton industry cost-effective conversion of
cellulose from waste plant parts into
ethanol*
• • A large amount of high-quality nanosilica is
produced from Rice Husk which can be
further utilized in making other materials such
as glass and concrete.
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185. Nanotechnology in Food
Processing and Packing
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216. Nanotechnology in Environment
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229. Impacts of nanotechnology
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230. Future of Nanotechnology
• As in biotechnology, issues of safety on health, biodiversity, and environment
along with appropriate regulation are raised on nanotechnology.
• However, nanotechnology products such as antibacterial dressings, stain-
resistant fabrics, and suntan lotions are available.
• Dream of automated, centrally controlled agriculture can become reality now.
• Modern agriculture is need of hour because conventional agriculture will not
be able to feed an ever increasing population with changing climate, depleting
resources and shrinking landscape.
 Experts says that nanotechnology will likely create the next generation of
billionaires and reshape global business.
 Industry Analysts Predict Revenues from Products Incorporating
Nanotechnology to Reach Close to $3 Trillion US Within 10 Years
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232. Implications of Nanotechnology
 Health and safety issues
 Nanoparticles can cause serious
illness or damage human body.
 Untraceable destructive weapons
of mass destruction.
 Social & Political issues
 Creates social strife through
increasing wealth gap
 Advisability of increasing scope
of the technology creates political
dilemma
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233. Nanoethics
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234. Health and Safety Issues
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• Great debate regarding to what extent nanotechnology will effect human
health
• Small nanoparticles may enter the human body but the health implications
are yet unknown
• Health effects can not be studied b/c all studies are made on animals not
humans
• So, difficulty in relating reactions to humans
• Toxicity studies using mice and rats suggest that certain nanomaterials could
be very toxic
• Safety in handling of nanoparticles
• Use of implanting nano-devices in humans: i.e. implanting artificial devices
Nanotechnology's health impact:
a. Nanomedicine; as medicine
b. Nanotoxicology; exposure to nanomaterials

235. Medical Issues
• Nanoparticles can be used as vehicles for efficient
drug delivery to heal, repair damages
• Nanomedicine could harm the human body rather
than healing it
• Particles such as toxins that can’t be seen or easily
controlled would enter the body
• The materials used for nano-medical technologies
may be toxic
• Transhumanists – changing human nature itself
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236. Environmental Issues
• Nanopollution generated by nanodevices could
be dangerous
• Might enter humans, causing unknown effects
• Whole life cycle needs to be evaluated for
assessing the health hazards of nanoparticles
• ‘Grey Goo’
• Chances of wiping out the entire biosphere by
self replicating nanorobots
• Release of nanoparticles which may harm the
environment
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237. Societal Issues
• Broader societal impacts and social challenges
• Military and terrorist uses - Unfortunately, as
with nuclear technology, it is far easier to
create destructive uses for nanotechnology
than constructive ones
• Fear of decrease of gap between humans and
robots
• Patent issues
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