Nano Biotech

7/27/2019 Nano Biotech

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nanobiotechnology

&bionanotechnology


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Intro

Two of most promising technologies of future:

Biotechnology: Use of living in the creation of wealth (products

or processes)

Nanotechnology: creation, investigation and utilisation of

systems that are 1000 times smaller than the componentscurrently used in the field of microelectronics.

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


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Intro Functional biological assemblies are inspiration fornanotechnological systems and devices

Molecular recognition btw. building blocks self-assemblyformation of functional devices

motors, pumps, cables, etc, all functioning at the nano-scale


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What we should know and what are the

possibilities??

Interaction between biological and non-biological devices???

Interactions with biological as well as non-biological substrates

Toxicity

How does nature make use of adhesive and anti-adhesive interactions?

Screening methods in biology

Bio-Chips

Lab-on-a-chip

Nanotechnologically modified biomaterials

Nano aspects of biological systems

Nanotechnological tools to improve biomaterials

Nanoparticles as therapeutic drug carriers and diagnostics

Drug, oligonucleotide, imaging agents

Nanodevices in medicine, pharmacy and biology


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Bionano-DNA as templateGazid E., FEBS Journal, 2006

DNA is very suitable for nanotechnological applications from thematerial science point of view:

1. The diameter of ssDNA is less than 1 nm

2. DNA molecules are chemically very robust

3. Low cost of large-scale chemical DNA synthesis

4. Easy modification: for example, by biotinylation or thiolation


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Bionano-DNA as templateGazid E., FEBS Journal, 2006

Examples:

DNA used in the formation of nanowires (1998): Metallization of

dsDNA btw two gold electrodes to form conductive silver nanowire

DNA-binding proteins (Figure)


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DNA Codes for Nanoscience

a) Holliday junction

b) Assembly of gold nanoparticles

c) Immobilization of gold NP

d) PCR mediated introduction of new

fuctionalities to create DNA-protein hybrids

e) Self-replication of connectivity


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Inspired by Nature-1Yusko, E.C et. al, Nature Nanotechnology, 6:253260, 2011

Challenges to reach the full potential of nanopore-based sensing:

reliable fabrication of synthetic nanopores on the sub-nanometre

scale

better control of translocation times of single-molecule analytes methods to control the surface chemistry inside synthetic pores:

reduce non-specific interactions of analytes with the pore walls and

prevent pore clogging

low frequency of translocation events at low analyte concentrations

and the poor specificity of the nanopores for analytes need to beimproved


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Fig 1: Insects detect pheromones by

translocating odorant molecules

through lipid-coated nanopores (D:

665 nm)

Fig 2: Lipid coatings are thought toparticipate in the capture, pre-

concentration and subsequent

translocation of odorants to specific

receptors

Fig 3: Capture, affinity-dependent

pre-concentration and translocation

of specific proteins after binding to

ligands on mobile lipid anchors


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Clogging Problem: Amyloidogenic peptides: e.g.

Alzheimer's disease-related amyloid-beta (A) peptides


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Self-Assembly of a Viral Molecular

Machine from Purified Protein and RNA

ConstituentsPoranen et al, Molecular Cell, Vol. 7, 845

854,

2001

Understanding of self-assembly innature


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Cellular imaging


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in Cell

Cell tracking: Different

population of cells in

tissue


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in Cell


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in Cell

Photo-thermal therapy


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in Cell

MRI and Cell Tracking

Fate of cells in the implantedarea

Anticancer therapy


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in Cell


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Nanotech in Drug Delivery

Controlled drug-delivery systems deliver drugs in the optimum

dosage for long periods

increasing the efficacy of the drug

maximizing patient comfort

enhancing the ability to use highly toxic, poorly soluble or relatively

unstable drugs

Nanoscale materials can be used as drug delivery vehicles to

develop highly selective and effective therapeutic and diagnostic

systems

Nano vs micro nanoscale particles can travel through the blood stream withoutsedimentation or blockage of the microvasculature

Small nanoparticles can circulate in the body and penetrate tissues

nanoparticles can be taken up by the cells through natural means such

as endocytosis


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Nanotech in Drug Delivery

Particle Size, Surface-to-Volume Ratio, Surface Area, and Surface Free

Energy

Biological Reactivity

Opsonisation: thought to be the greatest threat engulfment of foreign

particles injected into the blood stream by specific macrophages cells of

RES (reticulo endothelial system)

Modifications:

Nonadhesive surface coatings

Attachment of molecules for targetting

Layer-by-layer methods: shown to regulate nanoparticle biodistribution:cationic pegylated liposomes are preferantially uptaken by the liver and

tumor vessels in stead of spleen and blood accumulation

Synthesis from amphiphilic polymers


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Nano-Layered Microneedles for Transcutaneous Delivery of Polymer

Nanoparticles and Plasmid DNADeMuth et al, 2010, Advanced Materials

A) SEM micrograph of uncoated

PLGA microneedle arraysB) Polyelectrolyte layers

A) 24 bilayers for 5 min

B) 1 bilayer for 24 h

C) 5 bilayers for 24 h

D) 24 bilayers for 24 h

Luciferase gene and lipid-coated

PLGA NPs were delivered

seperately.


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Nanoparticles for ex vivo siRNA delivery to dendritic cells for cancer

vaccines: Programmed endosomal escape and dissociationAkita et al (2010) and Kogure et al (2007)J. Cont. Rel

Solution??

Programmed packaging


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Nanotech in Medicine: Oncology

It can complement existing technologies for detection,prevention, diagnosis and treatment

Useful in the area of biomarker research and increase

sensitivity in assays with relatively small sample volume

Jain, KK, BMC Medicine 2010, 8:83


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Nanotech in Tissue Engineering

For proper function and organization, we should

mimic native tissues at the nanoscale

Fabrication: top-down, bottom-up

Modification: Microfabrication and nanofabrication tomodify surface properties with resolutions as small as

50 nm control of cell behavior, orienting cells and

guiding cell migration, differentiation??


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Cell interactions with hierarchically structured nano-

patterned adhesive surfacesArnold, M, et al, Soft Matter, 2009, 5, 72

Counting the number of clustering cell adhesion based

transmembrane proteins is performed by molecular

defined, biofunctionalised nanopatterns of defined single

protein binding sites confined in micrometre large areas,

i.e. hierarchically organised micro-nanopattern


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Nanotech in Bio-Sensing


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Nanotech in Medicine: AMPs

Review: Calderon et al, Amino Acids (2011) 40:2949

Cationic nanoparticles formed by the conjugation of

cholesterol and antimicrobial peptides (AMPs): to

cross the bloodbrain barrier for treatment of fatal

Cryptococcal(Wang et al. Biomaterials 31(10):2874

2881 2010)

Nanostructured thin films with immobilized AMPs as

an agent intended to combat and prevent infection and

formation ofStaphylococcus biofilm related implantfailure (Shukla et al. Biomaterials 31(8):23482357,

2009)


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Interface: NSA-1

1. Park, S; Hamad-Schifferli, K, Current Opinion in Chemical Biology, 14: 616-

622, 2010

2. You, et al, Nano Today 2 (2007), 3443

3. Park and K. Hamad-Schifferli, ACS Nano 4 (2010), 25552560

The biological behavior of nanomaterials depends primarily on how

they interface to biomolecules and their surroundings

Issues like non-specific adsorption (NSA) are still the biggest

obstacles and have held back widespread practical use of

nanotechnology in biology


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Interface: NSA-2

Utilizing NSA:

(a) Tunable intracellular release from

NPDNA nanoplexes

(b) Enhancing protein translation: In

vitro gene expression with DNA,AuNP recruits mRNA and

translation related molecules into

its proximity

(c) Protein coronas induce a

biological response


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Nanonetworks

Communication???

Nanomechanical

Acoustical

Electromagnetic Chemical or Molecular

Short-range:

Molecular motors

Ca2+ signalling

Long-range:

Pheromones

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