Magnetic Bead Technology

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16 Νοε 2013 (πριν από 3 χρόνια και 8 μήνες)

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Magnetic Bead
Technology

History, Applications, and Marketability


University of Massachusetts
-
Lowell


2011

Shiv Sharma

Garo Yessayan

Zachary Nicoll

George Chahwan

Jason Tarantino


2


Abstract

The objective

of this review

is to track the on
-
going progress and evolution of the
biotechnological field through the experimentation with magnetic bead technology and its
applications to modern society.
The

review will

look at the history of magnetic bead technology,
from its development t
o its current applications, its

effect on the scientific community
, and its
marketability
.

Introduction

Background

Magnetic Bead Technology became popular in the 1980’s w
ith the discovery that
magnets can be used to more accurately perform already present technology. The technology is
based around the use of para
-
magnetic materials that act as magnets in the presence of a
magnetic field but are no longer magnetic when the
field is gone. This allows scientists to study
the materials effects in a magnetic field and then separate the proteins and peptides to get
physical results. Major companies have become interested in the development and application of
the technology so it
has been able to grow rapidly. The major upsides of this technology are that
it is significantly cheaper, requires less labor and is widely applicable. These advantages are the
driving forces behind the conception and development of the magnetic bead techn
ology. Some of
the applications are highly interesting and can be productive in the medical field. Bar
-
coded
magnetic bead (BMB) technology is new, upcoming and is currently being introduced to society.
This technology can help us in the medical field by a
ssisting doctors in the bacteria testing
department. Bacteria can be quickly identified and can also get accurately matched up with the
best possible antibody. This can cut days off the old methods of testing which can not only
improve a patient’s health

faster, but can also hinder anti
-
body resistance by picking the

3

strongest medication for the job. In addition, these advancements will offer new opportunities for
automated, low
-
cost and fast cancer diagnosis.

The Needs and Opportunities

Lately,

there has been a seemingly

eternal quest for improved medical care at lower costs.
With an aging US population In Vitro Diagnostic (IVD) companies are scrambling to find
biological solutions that will bring down the price of improved medical care. Due to
this urge,
the IVD industry is rapidly
evolving
. In a nutshell, the need for improved care at a low cost
entails improved diagnostic methods. Therefore, a lot of recent research is focused in figuring
out more
efficient

and precise
diagnostic methods. An e
xample
would include improved testing
sensitivity
. Improvements in this
phase of the diagnostic method will lead to better diagnosis, a better clinical management, and
inevitably reduced costs of clinical care.
Thus, IVD companies have been looking at nume
rous
technologies such as magnetic beads to help if finding a solution to the needs of consumers.

Magnetic beads, in the last twenty years have exponentially progressed to now be
considered the “golden standard” in the magnetic microspheres and magnetic na
nospheres field.
Magnetic beads are an essential component of immunoassay diagnostic kits for analyzers in
clinical labs. These microspheres are actually polymer shells with a magnetic pigment; here, the
magnetic material is mainly iron oxide. Functional g
roups of the polymer surface permit
chemical derivatization of magnetic particles. This process allows the conversion of magnetic
particles into a binding agent for tests using immunoassays.

In the biomedical field, magnetic beads range in size anywhere b
etween 1um
-
100um,
being 1um
-
2um in size on average. The iron content of these beads also varies between 15%
-

4

60%; this amount determines the response to an applied magnetic field, facilitating the
manipulation of an
y phase in any biomedical test.

Instrument
ation

Turbo Beads


Turbo Beads are the next generation of magnetic beads; they are magnetic nanobeads that
have a core and shell structure. The beads core is constructed of metal, and its shell is made out
of carbon. The metal core is used because of its
highly
magnetic properties, and the carbon is used to provide
a chemical stability. This new technology provides a
fast and efficient way to separate various compound
from one another.


This technology began by taking highly
reactive metal nanomagnetics a
nd coating them in graphene
-

carbon. The binding of the beads is
a carbon to carbon bond. This covalent bond allows for no ligands to be lost. A ligand is an atom
that is bonded to the central metal atom. The graphene
-
carbon allows the beads to have high
thermal stability. This means that the beads can be used in areas that had low pH levels (Acids)
and high temperatures without oxidation of the core.


That is not the only positive property of the
turbo bead. Due to the combination of the metal
core and c
arbon shell these beads display a large
increase in their magnetic properties. Below is an
image that demonstrates how much more reactive
the turbo bead is compared to a ferrite based bead.


5

In
the

image
on the left
we can see that the ferrite based bead

is located to the left and is in a red
color; whereas the turbo bead is located to the right and is in a black
color. In the middle of the two tubes is a standard magnet. It is clear
that after forty seconds the turbo bead concentration is comparatively
m
uch greater than the ferrite bead concentration.


The picture
on the side

is another example demonstrating how
quickly the turbo beads can separate from a substance. In this image
there is a substance that contains the turbo beads. It can be seen that
once

a magnet is applied to the vessel (bottom left corner) it only takes five minutes for the turbo
beads to completely separate from the substance.

The chemical properties of the turbo beads allow them to have a selective separation of
precious metals at
low concentrations. Also, their highly magnetic properties allow for high
recyclability of magnetic chemicals, which permits them to be reused.


Today these turbo beads are being used for toxin management in water streams. Given
that the beads are highly

reactive and chemically stable, they can be used for the selective
separation of heavy metals. The compounds that create the magnetic bead result in reagents for
swift removal of toxins that can be found in
contaminated water.

The picture
on the left

is

a graph that shows
the amount of cadmium that is in water before
and after the use of turbo beads. It can be
seen that using the turbo beads significantly
decrease the amount of cadmium found in

6

contaminated water. This means that turbo beads allow the pu
rification to go down to drinking
water standards.


CardioGenics Magnetic Beads

CardioGenics
’ bead

is
a type of magnetic bead that was developed to improve testing
sensitivity. Most commercial magnetic beads have a dark color to them, and approximately 80
%
of
the light they generate is lost, causing

them to have a low testing sensitivity.


The CardioGenics magnetic bead is a lighter colored bead that is used to optimize and
collect light signals in binding tests. These beads are a lighter color than the ge
neric bead
because they are coated with a thin layer of silver before they are covered with a polymer shell.
Since these beads are lighter they become more sensitive to light; which in turn maximizes light
collection. The CardioGenics

can vary in size f
rom 1 to 50 microns, and
have seven times more
light sensitivity than a generic magnetic bead.
Due to the increase in light sensitivity these beads
improve the testing sensitivity.


Below are two images that show the different amounts of silver coating that can be added
in order to change the color of the beads. The lighter the beads are the more sensitive they are to
light.






7

Asynchronous Magnetic Beads

A

new promising technolog
y is the asynchronous magnetic bead rotation (AMBR).

In
order for this technology to be useful in the field of pharmaceuticals and medicine,
the beads
need to be extremely sensitive as the beads
are to examine various biomolecules on the
micro
even nano
scale.


This breakthrough technology was
elucidated through the m
onitoring
of the
continuous growth of individual bacteria by
direct observation using optical imaging.
Unfortunately, direct observation has
numerous

limitations such as o
ptical diffraction

and a limit to the

number of cells
that
co
uld be
continuously monitored. On the brighter side
, advances have been made to increase the rotational
speed of the AMBR to 145 Hz which allows
the bead to have
a detection limit of 59nm. Such
precision could be used to measure nanoscale growth dynamics of individual bacterial cells.

AMBR sensors operating at
high frequencies of 145 Hz allow

for
higher resolution
candid; this

is important
while

dealing with molec
ules on a very small scale. Even though
other
technologies like electron microscopy and
cantilevers already o
ffer high resolution
monitoring, t
he drawback of using electron
microscopy and cantilevers is that they give
optimal results in vacuum or air. AMBR

sensors
will make real time single bacterium growth

8

monitoring and single virus detection possible in their given fluid environment.













A rotating magnetic field controls the rotation of a ferromagnetic bead. The bead rotation
becomes asynchronous with the magnetic field at a critical frequency Ωc. The critical frequency
depends on the magnetic momentum of the bead
m,

the magnetic field st
rength
B
, the kinetic
viscosity
ɳ
, the volume of the bead
V
, and the shape factor k (for a sphere, the shape factor is 6)

Applications

Turbo Beads/

CardioGenics Magnetic Beads

Applications

With magnetic beads being such a new but fast growing technology,
there is no doubt
that this technology can be applied to numerous fields

shown in the figure below
. A few fields
include clinical diagnostics, drug targeting, cell isolation, purification, nucleic acid purification
and detection.




A very popular applica
tion of magnetic beads would be providing more efficient assay
platforms compared to suspension bead based assay platforms. Immunoassays are used to
investigate the roles of biomolecules. Immunoassays help with trying to stop the progression of
diseases su
ch as HIV, Alzheimer’s disease, and numerous cancers. Immunoassays can use both

9

magnetic bead platforms or
suspension platforms but there are
numerous reasons as to why magnetic
beads are a better choice.


At a glance, there are various
drawbacks to suspe
nsion beads.
Though these beads offer numerous
advantages in the areas of sensitivity
and cost, there are many unintended
consequences that are elucidated
during the process of use.
Examples
include the vacuum that is used to
wash and remove liquid from th
e
microscopic beads cause pressure that
can fluctuate between wells and
plates. This can ultimately skew data
results. Also, human error is a major
influence on the results in suspension
bead tests are a lot of manual care
-
take is involved.


On the contr
ary, magnetic beads just make the whole testing process more efficient and
precise. The magnetic bead based assay platforms use a series of magnetic beads that have
different dyes and emit at varied wavelengths. This is done to create a unique spectral add
ress for

10

each bead. These beads then serve as solid phases to capture analytes. The figure on the side
describes the process of the magnetic beads in the assay platform.


Using magnetic beads to
carry out the process of magnetic separation in an automated

wash eliminates the variables that cause skewed answers with suspension bead use. Additionally,
with magnetic bead technology
,

assays can be completed in 3
-
4 minutes and do not require
additional user training.
Conclusively, the benefits of using magnetic

beads outweigh those of
suspension beads.

Magtration Technology

Magtration technology in simple terms is the filtration for a nucleic solution by means of
a magnetic force. When studying these nucleic acids, one can learn much about how cells
function, research diseases, discover new medicines, and countless more appli
cations. Nucleic
acids are known as the building blocks of life and contain different types of DNA and RNA.
These are found inside every animal cell and we can now separate nucleic acids by implementing
magtration technology.

Every cell is surrounded b
y a cell membrane which must be broken down first in order to
get to the inside of the cell where nucleic acids are found. To do this, there are certain toxins and
detergents that will break down a cell membrane but not harm the internal parts of the cell
. A
separate solution containing these paramagnetic beads is added to the lysis solution. These
paramagnetic beads are coated with silica with is a natural attraction for the nucleic acid. This
new mixture is shot up and down a pipette in which the magnet
ic beads are trapped up against
the wall where a magnet is placed. Now that the magnetic beads and the nucleic acid trapped in
the pipette, it must go through separate washing solutions. Several wash cycles should be
preformed to maximize the purification

process. From here, the beads are moved into a new
container containing a low ionic strength solution. When this solution is warmed up, it separates

11

the beads from the nucleic acid. Then the final step is to remove the beads from the solution by
use of

a magnet and aspirating the solution up and down to collect the beads as done earlier and
what remains is the purified nucleic acid.

Asynchronous Magnetic Bead Applications

As
mentioned earlier, researchers are looking to operate
AMB’s
at a high frequency
because that would mean a higher resolution and a higher sensitivity
caused by changes

in
diameter. Many current applications would benefit from a higher critical frequency;

examples
include

micro
-
mixing and growth studies. Using a ferr
omagnetic bead to reach a bead frequency
of 145 Hz (shown in graph 1) that al
lows a detection limit of 59 nm; this is sen
sitive enough to
study single bacterial growth.



















Graph 1 (1)


The rotational frequency of the bead is calculated using the following formula:























Many applications

use the sensors to monitor the growth of individual bacteria
. In a
recent experiment the sensors were used to monitor the growth of bacteria

and its susce
ptibility

12

to a particular drug.
The Escherichia coli also known as E coli
bacteria along with
the
ant
ibacterial ampicillin

were used for the experiment

(2).

The experiment showed that the AMBR sensor was able to detect changes

at even 80nm
in a single
E coli

cell. The sensor was

also
able to detect the response of the
E coli
cell to the
antibacterial
used. What is also worth mentioning is the fact that the experiment was done in
water
; this

shows that the sensor is able to work in an aqueous environment unlike previous
technologies like electron microscopy that would work best in a vacuum environment.
The
success of this experiment has extended the method to similar single cell studies like cancer
cells.

Finally, we can conclude that asynchronous magnetic bead rotation technology has a lot
of potential. It has the ability to speed up treatment of bacter
ial infections by allowing us to find
antimicrobials for infections in minutes instead of days. Another field where the technology
could be of vital use would be for studying the response of individual cancer cells; such tests
could equal huge advances in
drug development and treatment. The AMBR technology will help
get faster results at a cheaper price hence saving more lives.


13


Radiation Checking Magnetic Beads

Professor Lawrie Challis, chairman of Mobile telecommunications and health and
research program
me, has been looking into the affects of mobile phone radiation and
alternatives. Mobile phones output electromagnetic
radiation which
, at
10^(
-
8) Hertz

resem
ble the
frequency of microwaves
.

C
ell phones
are not the
only technology to emit such radiation;
in fact

any
type of wireless communication device will use the
same technology. Excessive use of these devices can
lead to many health risks. Two of the leading problems are developing brain tumors, and break
down of
the

blood brain barrier.
The

blood b
rain barrier protects the brain from incurring
harmful substances that are within the body. When it breaks down it makes your brain
susceptible to many health dangers. Two leading diseases that are related to the BBB breakdown
include Alzheimer’s disease

and Parkinson’s disease. Other smaller risks include constant
headaches, sleep disturbances, memory loss, learning disabilities, and infertility.


Every device has a specific absorption rate which is the measure of the amount of radio
frequency energy

is absorbed by one’s body when
using these device headsets which determine the
danger of the device. All these risks decrease
vastly with increased distance from the device to
your body. This is why hands free devices are
very good
based on the

fact tha
t they cut down

14

immensely the amount of radiation that is absorbed into your body. Unfortunately
,

they do not
eliminate these harmful risks.


Professor Challis has done much research and found that attaching a magnetic bead to the
wire or antenna of the
phone. This type of bead is called a ferrite bead because it acts as a
suppressant to the hig
h frequency noise in the device;

i
t stops any current from passing it on the
outside of the wire. In simple terms it means that no radiation is pulsing out of th
e phone and
is
unable to

reach the head. This technology can be used as a marketing technique that will
promote certain phones that contain the beads as “health safe phones”.

There is ongoing debate on whether these magnetic beads are a necessity within t
he cell phone
unit. There are two main arguments against the use o
f magnetic beads in cell phones: o
ne is that
the cell phones pass a government safety test which regulates the amount of radiation a
phone
can legally have;

t
he United States requires a 1.
6 watt per kilogram SAR rating (specific
absorption rate)
. The other argument against the use of magnetic beads in cell phones

is that
since the effect of the radiation takes a very long time to develop, not all scientists are convinced
that there is a li
nk between the radiation and
any illnesses. The scientists argue that the
se
illnesses can be caused by many effects and not just radiation poisoning.

Illness Detection

Magnetic bead technology has also been linked with the purification and detection of
timorous cells. Mixing magnetic bead technology and micro
-
electro
-
mechanicals
-
systems
(MEMS) has resulted in the ability to detect tumor cells rapidly and it also has the
capability of
collecting and purifying them. The magnetic beads allow specific antibodies to be incorporated
onto their surface to target and recognize the desired tumor cells in the clinical body fluids. The
whole idea is based around the technology devel
oped called a 3D micro
-
incubator. The machine

15

itself was developed for the incubation and mixing process, and it has the ability, using the
vortex effect, to mix large amounts of bio
-
samples efficiently and rapidly. It has also
successfully detected ovaria
n and lung cancer cells
by performing on
-
chip identification. The
resulting
mixture of magnetic beads and the 3D micro
-
incubator leads to a new ability for rapid
purification and detection of cancer cells.


The need for a means of detecting cancerous
cel
ls has long been a top priority in the scientific field since there exists no known guaranteed
cure. Currently we are capable of detecting cancerous cells using biomarkers of tumor cells, cell
biopsy and magnetic resonance imaging (MRI), but these methods
of detection take a long time
and require complicated machinery and procedures. Therefore the need for a rapid means of
detection exists. It is known that an early detection of cancerous cells is crucial to the prevention
of the cells metastasizing, so a t
echnology enabling
very early detection is highly sought after. With the
emergence of MEMS technology we have been
able to miniaturize biomedical devices and
systems. The combination of MEMS technology
and its use in Biology has led to the emergence of a
n
ew field called lab
-
on
-
a
-
chip technology (LOC).
This technology means that a whole host of procedures and devices can be placed onto a single
microchip, and it also reduces the amount of time it takes to diagnose because the process of
analysis and prepara
tion have been reduced to


an automatic function. Using these LOC devices,

16

scientists have been able to perform cancer analysis using cell sorting, separation and
spectroscopy techniques. This technology may possibly lead to the ability to specify treatmen
ts
to individuals, however the process still requires lots of bulk equipment that are needed to
perform the identification of the cancer cells. This is where the implementation of magnetic
beads comes in to play. We are able to surface
-
modify magnetic bead
s, which allows for a
specific type of cell to be detected rapidly, and when combined with the previous technologies
the overall process becomes much quicker.


An actual experiment for the purification and detection of tumor cells was performed to
accurat
ely test the hypothesized outcomes. First they extracted body fluid in large amounts,
around 20mL. The body fluids were then pre
-
centrifuged and collected. The next step involved
re
-
suspending them with phosphate buffered saline into a 1 mL volume in an ep
pendorf tube.
The samples were then incubated with magnetic beads specifically coded to recognize the tumor
cells which could then be purified by using a magnetic field. The actual purification process is
performed by using a vacuum pump to suck out all of

the interference substances, leaving only
the magnetized beads behind. The target mRNA can then be reverse transcripted and amplified
using a built
-
in
-
self
-
compensated temperature control module.


The overall layout of the system is comprised of three m
ain units; the 3D micro
-
incubator, a microfluidic control module and a nucleic acid amplification module. The 3D micro
-
incubator has three layers of PDMS structures which allow large quantities of fluids to be mixed
quickly. Each layer has a thick PDMS str
ucture and a thin pneumatically
-
driven PDMS
membrane which generates the mixing effect. The thick PDMS structure is made up of two air
chambers with a connecting chamber. It has a built in electromagnetic valve capable of driving it
via a digital controlle
r and a vacuum pump. The vacuum pump generates the vortex flow in the

17

mixing chamber, which is what is needed for the incubation process. For final proof, two kinds
of cancer cells were incubated with magnetic beads, ovarian cancer and lung cancer. The
mag
netic beads were conjugated with the correct antibodies and placed in the incubator. Using a
magnetic the beads were attracted and the unwanted substances were sucked out with a vacuum.
It was recorded that almost all of the cancer cells were captured and
purified by the experiment.
On average about 92% of tumor cells are capable of being specifically targeted by this
technology. The use of this technology will hopefully bring about the ability to detect and
prevent cancer, that way we can avoid the need to

cure it.


Marketability


Magnetic beads are the most
widely used solid phase for
automated
methods

for isolation
and detection of biomolecu
l
es. In
fact, 9/10 top IVD companies use
magnetic beads in their automated
analyzers. Additionally, binding
tests (such as immunoassays and molecular tests) combine for 1/3 of the clinical tests done in the
market. These tests use magnetic beads as the primary component. This market is also value
d at
USD $42 billion as of 2008. Magnetic beads
themselves have a market of
$1 billion for
immunoassay and molecular diagnosis.


Furthermore, Dynal, a leading magnetic bead manufacturer claims that while the IVD
market is fast growing, the magnetic beads m
arket is growing exponentially faster. In the same
report, Dynal explains that immunoassays make up $4 billion if the IVD market where magnetic

18

beads
are once again the golden
standard. Moreover, nucleic acid testing
makes a smaller portion, roughly $2
bi
llion of the IVD market; here,
magnetic beads are the most common
solid phase employed. Even areas such
as genomics that include DNA and RNA extraction and purification (a $2.3 billion market) have
started using magnetic beads at a growing pace.

Conclusio
n


There is no doubt that magnetic bead technology is the most modern and progressive
technology available in the market today. Being in the market for only a few years, magnetic
beads have gone from niche experimentation to a staple in the biomedical fiel
d. With its
numerous practical applications such as asynchronous bead rotation, radiation sensing abilities
and biomolecules separating abilities, magnetic beads are a technology that will be implemented
for numerous years to come. Even from a marketing po
int of view, the technology has a lot of
room to gro
w in the market; based on a simple cost analysis it can be seen that this technology
will only rise monetarily for the fore coming future.









19

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-
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