磁珠在臨床以與生物科學(xué)和技術(shù)中運(yùn)用

Magnetic

nano-

andmicroparticles

in

bioscienceand

biotechnologyIvo

Safarik

and

Mirka

SafarikovaCeske

Budejovice,

Czech

RepublicType

of

magnetic

nano-

andmicroparticlesMulti

domain,

single

domain

orsuperparamagneticMagnetite

(Fe3O4)Ferrites

(MeO

.

Fe2O3;

Me

=

Ni,

Co,

Mg,

Zn,

Mn)Maghemite

(

-Fe2O3)Greigite

(Fe3S4)Iron,

nickelMagneticnano-

andmicroparticlesInorganic

material,

but:Present

in

biological

systemsUsable

for

a

variety

of

bio-applicationsImportant

dates

1962

-

Lowenstam

(biochemically-precipitatedmagnetite

as

a

capping

material

in

the

radula(tongue

plate)

teeth

of

chitons

(marine

mollusksthe

class

Polyplacophora)1975-Blakemore

(magnetotactic

bacteria)Magnetic

material

in

teeth

ofchitons

hardening

the

toothcaps

(enabling

thechitons

to

extract

andeat

endolithic

algaefrom

within

the

outerfew

millimeters

ofrock

substrates)Magnetotactic

bacteria

Various

morphologicaltypes

exist

Gram-negativeprokaryotes

Magnetic

nanoparticlespresent

in

magnetosomes

Found

in

sediments

indiverse

aquaticenvironmentsMagnetotactic

bacteriaProvidedby

Dr.

D.

Schüler,Magnetosomes

Composed

from

magnetite

(Fe3O4)

or

greigite(Fe3S4)Ca

50

nm

in

diameterCoveredwith

a

lipidic

membrane

Important

for

navigation

of

the

bacteria

intooxic/anoxic

layerBiocompatible

character

(phospholipid

bilayerPossible

applications

in

biosciencesMagnetosomesProvidedby

Dr.

D.

Schüler,

GermanyMagnetosomesProvidedby

Dr.

D.

Schüler,

GermanyBiogenic

magnetic

nanoparticlesMagnetotactic

bacteriaMagnetotactic

algaeProtozoaInsectFishAmphibiansReptilesBirdsMammalsAnimal

magnetite

Magnetosome

chainsfrom

the

frontaltissues

of

chinooksalmonS.

Mann

et

al.:

J.

Exp.

Biol.

140(1988)

35-49.Importance

of

animal

magnetite

Most

probably

involved

inmagnetoreception

(what

is

the

currentposition

???,

which

direction

to

move

???interaction

of

magnetite

nanoparticles(chains

of

magnetic

nanoparticles

?)

withthe

sensing

cellsNon-physiological

presence

ofmagnetic

nanoparticles Found

in

human

brain

in

case

ofneurodegenerative

diseases

Disruption

of

normal

iron

homeostasis

inthe

brain

Early

detection

of

neurodegenerativediseases

using

MRI

??Magnetic

nanoparticles

inmeteorites

from

Mars

Meteorite

magneticnanoparticles

resemblemagnetosomes

fromEarth

magnetotacticbacteria

Hypothesis:

Was

therea

transfer

of

livingsystems

from

Mars

toEarth???Let’s

return

back

to

Earth

!!Magnetic

nano-

and

microstructuredmaterials

for

bioapplications

Useful

for

many

applications

in

bioscienbiotechnologies

and

environmentaltechnologiesEnable

to

label

diamagnetic

structures

Enable

separation

or

detection

ofmagnetically

labeled

structures

Both

separation

and

non-separationapplications

are

importantWhich

nano-

and

microstructuredmagnetic

materials?Three

important

bio-

Biocompatible

(in

the

broadest

sense

of

tword)BiodegradableBiomimeticBiocompatible

materialsBiocompatibility

in

the

broadest

sense

of

the

wordMinimal

toxicityStability

of

nanoparticles

in

body

fluids

Possibility

for

immobilization

of

biologically

activecompounds,

cell

organelles,

cells

(activated

nanopartReproducible

interaction

with

biosystemsEnable

labeling

of

biostructures

(for

magnetoassays)

Biopolymer

based

particles,

PEGylated

structures,magnetoliposomes,

biopolymer-based

FFs,

self-assembmagnetic

structuresBiodegradable

materials

Important

from

the

point

of

view

of

possibclinical

applications

(e.g.,

drug

target

Nanoparticles

formed

by

biodegradable(bio)polymerMagnetoliposomes

and

related

structuresStarch-based

magnetic

fluidsBiomimetic

materialsAnalogues

of

bacterial

magnetosomesFerritin

x

magnetoferritinImportant

data

1960s

–

Pappell

(first

ferrofluids,

1000

hoursmagnetite

milling)

1966

-

Magnetically

controlled

thrombosis

ofintracranial

aneurysms

1970s

–

increase

of

interest

in

biomagneticresearchandtechnology

1980

–

Massart

(chemical

synthesis

of

aqueousferrofluid

without

detergents)1980s

–

commercially

available

magnetic

particMagneticfluids

–

examples

of

magneticnanostructured

materials

ferrofluid

can

beinfluenced

by

thepresence

of

externalmagnetic

fieldSynthetic

biocompatible

magnetic

nanoparticlesandcomplexescontaining

them

Biocompatible

magneticfluidsMagnetoliposomes

Magnetic

nano-

andmicroparticlesMagnetoferritinMagneticmicroparticlesDynabeads

Homogeneous

sizedistribution

Simple

recovery

withsmall

permanentmagnets

Magneticnanoparticles

presentinside

the

beads

Available

in

plain,activatedand

bioactiveExamples

of

of

magnetic

nano-and

microparticles

applications

From

molecular

biology

to

environmentaltechnologies

Manipulation

of

microliters

as

well

asmillion

of

litersManipulation

in

suspension

systems

Both

separation

and

non-separationtechniques

are

importantImmobilization

and

modificationof

biologically

active

compoundsPossibility

of

simple

removal

of

the

immobilizedmolecules

or

cells

from

the

system

Immobilization

of

enzymes,

antibodies,

lectinsoligonucleotides,

affinity

ligandsModification

of

proteins

via

magnetic

PEGImmobilization

in

magnetoliposomes

Magnetosomes

from

genetically

engineeredmagnetotactic

bacteria

carrying

target

proteinsIsolation

of

biologically

activecompoundsDirect

separation

of

targets

from

crude

samples

containing

solidsAffinity

magnetic

beads

or

magnetic

two-phase

systems

Preconcentration

of

target

molecules

from

large

volumes

of

crudematerialsPoly

A+

mRNA,

DNA,

oligonucleotidesEnzymesLectinsDNA/RNA

binding

proteinsAntibodies,

antigensXenobiotics

(both

organic

and

inorganic)High

gradient

magnetic

separatorsTest

tube

magnetic

separatorsAutomated

systemsKingFisher

mLThermo,

USAKingFisher

96Thermo,

USATe-MagSTecan,

JapanDetermination

and

detection

of

biologicallyactive

compounds

and

xenobiotics

Immunomagnetic

assays

(magnetic

particles

usedfor

separation

of

target

analyte)

Magnetoimmunoassays

(magnetic

particles

serveas

detectable

labels)Magnetic

substratesMagnetic

solid-phase

extraction

New

principles

for

the

detection

and

quantitatioof

both

magnetic

labels

and

biological

structuretested

(giant

magnetoresistance,

magnetoelastimagnetic

atomic

force

microscopy…)Automated

systems

forimmunoassaysntey

Combination

of

specificmagnetic

separation

andsensitive

detection

(enzyme,radioactive,

chemiluminesceor

fluorescent)

Different

assay

formats

can

bused

(sandwich

x

competitiveimmunoassays)

Generic

solid

phases

can

beused

(streptavidin,

secondarantibody,

protein

A

…)Beckman

Coulter"sAccessImmunoassaySystemMagnetoimmunoassays

Magnetic

nanoparticlesserve

as

a

label

Possibility

to

replaceenzymes,

radioisotopes

…

Magnetic

PermeabilityReader

(Euris,

Sweden,top;

Magnetic

AssayReader

(Quantum

design,USA,

bottom)Magnetic

solid-phase

extractionDetection

of

crystal

violet?inwater

Magnetite

withimmobilizedphthalocyanine

dye

usedas

a

selective

adsorbent

Curve

B

–

0.5

μg

of

dye

in1000

ml

of

water

Sensitivity

0.5

ppbachievedCells

Immunomagnetic

separation

of

target

cells

and

cellorganellesDetection

and

separation

of

cancer

cellsIsolation

of

stem

cellsSeparation

of

microbial

pathogensStudy

of

cell

functionTransfection

of

cellsMagnetic

twisting

cytometryMagnetic

labeling

of

cellsMRI

tracking

of

transplanted

cells

(magnet.

modified)Labeling

of

T-lymphocytes

withmagnetic

nano-

and

microparticlesE.

coli

O157

bound

to

DynabeadsPrinciple

of

IMSLarge-scale

immunomagneticseparation

of

cells

Several

magneticseparation

devicescommercially

available

CliniMACS

employs

magnetic

nanoparticles

forcells

labeling

and

HGMSsystem

for

separationMagnetotactic

bacteria

Biotechnology

production

of

native

orgenetically

engineered

magnetosomesNon-destructive

domain

analysisRadionuclide

recoveryDrug

and

radionuclide

targeting

Accumulation

of

active

compounds

in

thetarget

organ

or

tumor

Ferrofluids,

magnetoliposomes,

ferroflumodified

erythrocytes,

magnetic

polymercompositesMagnetic

drug

targeting

MTC

-

composite

of

elemental

iron

and

activated

carbonwith

anticancer

drug

adsorbed

onto

it

(FeRx

Inc.,

SanDiego,

CA,

USA)MagneticFieldBody

surfaceMagnetic

fluid

hyperthermiaCancer

treatment

Heating

of

tissues

labeled

with

magneticparticles

to

42

-

46

oC

after

exposure

toalternating

magnetic

field

Magnetic

fluids,

magnetoliposomes,magnetic

single-

or

multi-domain

particlCombination

with

drug

targetingSketch

of

the

first

prototype

MFH

therapy

system

(MFHHyperthermiesysteme

GmbH,

Berlin,

Germany)

–

Prof.

JordanContrast

increasing

during

MRISuperparamagnetic

contrast

agents

Biocompatible

ferrofluids,magnetoliposomes

Antibody-conjugated

magnetitenanoparticlesMRI

of

bone

marrow

PEGylatedmagnetoliposomes

asa

bonemarrow-seeking

MR

contrastagentPotential

medical

applicationsTreatment

of

AIDSEndocytosis

of

ferrofluids

by

cancer

celBrain

tumor

sensingArtificial

sphincter

muscleMagnetic

polymers

(artificial

muscle?)Environmental

chemistry

andtechnology

Waste

water

and

potable

water

purification

usingmagnetite

microparticles

(Sirofloc

process

–Australia)

Magnetically

modified

microbial

cells

forxenobiotics

removal

Magnetic

solid-phase

extraction

for

thepreconcentration

of

the

target

analytes

Magnetically

modified

enzymes

for

the

detectionof

their

inhibitors

(heavy

metals,

pesticides

etScheme

of

Sirofloc

ProcessInhibition

of

magneticallymodified

trypsinDependence

of

the

inhibition

of

thetrypsin

activity

on

theconcentration

of

Ag+

(●)

and

Pb2+(○).Dependence

of

the

inhibition

of

thetrypsin

activity

on

theconcentration

of

thionin

(

),

4-aminobenzamidin

(

),

safranin

(○)Adsorption

of

magneticnanoparticles

on

yeast

cells

Interaction

of

Saccharomycescerevisiae

cells

(bakers

yeast,brewers

yeast)

with

water-based

ferrofluids

Magnetically

modified

yeastcells

canbeused

as

anadsorbent

for

the

separation

ofbiologically

active

compoundsand

xenobiotics

(both

organicand

inorganic)

Adsorption

of

organic

dyes

ispresentedDyeQ(mg

ml-1

)Q’(mg

g-1

)Acridineorange13.182.8Aniline

blue68.1430.2Crystal

violet13.685.9Malachitegreen3.119.6Safranine

O14.390.3Future

trends

Biotechnology