APPLIED PHYSICS LETTERS VOLUME 85, NUMBER 14 4 OCTOBER 2004

Biological sensors based on Brownian relaxation of magnetic nanoparticles

S. H. Chung,a) A. Hoffmann, and S. D. Bader
Material Science Division, Argonne National Laboratory, Argonne, Illinois 60439
C. Liu, B. Kay, L. Makowski, and L. Chen
Bioscience Division, Argonne National Laboratory, Argonne, Illinois 60439
(Received 5 March 2004; accepted 13 July 2004)
We experimentally demonstrate a biomagnetic sensor scheme based on Brownian relaxation of
magnetic nanoparticles suspended in liquids. The characteristic time scale of the Brownian
relaxation can be determined directly by ac susceptibility measurements as a function of frequency.
The peak in the imaginary part of the ac susceptibility shifts to lower frequencies upon binding the
target molecules to the magnetic nanoparticles. The frequency shift is consistent with an increase of
the hydrodynamic radius corresponding to the size of the target molecule. © 2004 American
Institute of Physics. [DOI: 10.1063/1.1801687]

The biological and medical applications of micron or ␹ 0␻ ␶

nanometer sized magnetic nanoparticles have attracted recent
interest due to the successful development of various fabri-
cation techniques that combine fine magnetic particles with
biologically relevant coatings.1 This coating can have a bio-
logical recognition function for specific target molecules
through ligand–receptor and antigen–antibody bindings.
Since magnetic nanoparticles can be manipulated and de-
tected by magnetic interactions, biomagnetic nanoparticles
have been used in applications such as magnetic separation,
targeted drug delivery, or contrast enhancement of magnetic
resonance imaging. In addition, many recent efforts are fo-
cused on developing magnet based biodetection platforms.
Many of these sensor schemes rely on detecting the stray
magnetic field of the biomagnetic particles upon binding
them to a magnetic-field sensor using the target as a link.
These magnetic field sensors are based on anisotropic
magnetoresistance,2 Hall effect,3 or spin valves.4,5 Alterna-
tively, using a superconducting quantum interference device
(SQUID), biological binding activity was detected through
the relatively slow magnetic Néel relaxation upon immobili-
zation of the biomagnetic particles.6 One weakness of all of
the above sensing schemes is that they do not permit dis-
crimination between several different targets that may have
similar biological binding affinity. In this letter, we experi-
mentally demonstrate a substrate-free magnetic sensor
scheme based on the changes of dynamic magnetic proper-
ties of magnetic nanoparticles suspended in liquids. The ad-
vantage is that it allows distinguishing between several pos-
sible targets with similar binding affinity. Furthermore, it
allows for an inherent integrity check of the sensor, since a
useful signal is detected even without a target present.
The sensor scheme employed is based on the detection
of dynamic magnetic properties, as proposed theoretically by
Connolly and St. Pierre.7 The magnetic response of nanopar-
ticles suspended in a liquid to a small alternating magnetic
field with frequency ␻ can be expressed by a complex mag-
netic susceptibility ␹. The imaginary part ␹ corresponding
to the out-of-phase response is given by
⬙ Therefore, there is an upper limit on the magnetic particle
a)
Electronic mail: chungsh@anl.gov
␹ 共␻兲 = 共1兲
⬙ 1 + 共␻␶兲2
,
where ␹0 is the dc magnetic susceptibility and ␶ is the effec-
tive magnetic relaxation time of the nanoparticles in the liq-
uid. Note that ␹ peaks for ␻ = ␶−1. The magnetization of
⬙
magnetic nanoparticles suspended in a liquid can relax either
through a Brownian or through a Néel relaxation
mechanism.8 When the magnetic anisotropy energy is high
enough to block the magnetization inside the nanoparticle,
then the relaxation occurs due to rotational diffusion (Brown-
ian). However, if the magnetization is unblocked, then the
particles are superparamagnetic and the magnetization re-
laxes internally (Néel). Generally, the effective relaxation is
a combination of both relaxation mechanisms. For the case
that Brownian rotational diffusion is the dominant relaxation
mechanism, the relaxation time is
4 ␲ r 3␩
␶= , 共2兲
k BT
where ␩ is the dynamic viscosity of the liquid, r is the hy-
drodynamic radius of the biomagnetic nanoparticle, and T is
temperature. Thus, the peak in the imaginary part of the mag-
netic susceptibility is inversely proportional to the effective
volume of the nanoparticle. Therefore, if the hydrodynamic
radius increases due to the binding of target molecules to the
biomagnetic nanoparticles the Brownian relaxation time in-
creases and the frequency for the peak of ac susceptibility
decreases.
The dominant relaxation of the two mechanisms depends
on the particle size. Néel relaxation is the dominant mecha-
nism for particles less than 10– 20 nm while the Brownian
mechanism is dominant for larger particle diameters.7 How-
ever, when the particle diameter increases further the mag-
netization ceases to be single domain and a multidomain
state develops in order to reduce the magnetostatic energy. In
this case the magnetic relaxation no longer reflects the
Brownian motion, but instead can be dominated by internal
changes of the magnetization, i.e., domain-wall motion.
size for the sensing scheme presented here. Furthermore, be-
yond a certain limit the particles are not easily suspended in
a liquid. In fact, the use of small magnetic nanoparticles may

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2972 Appl. Phys. Lett., Vol. 85, No. 14, 4 October 2004 Chung et al.

FIG. 1. Particle diameters for stable single domain magnetic nanoparticles.

be beneficial in general, since they may avoid structural FIG. 2. Imaginary part of the ac magnetic susceptibility as a function of
change of the biological entities or blockage of the biological frequency for an avidin-coated magnetic particle suspended in a PBS buffer
binding interactions.4 Figure 1 shows suitable particle diam- solution. Solid symbols are measured at 300 K, open symbols at 250 K,
eters for the creation of stable single domain structures for which is below the freezing point of the carrier liquid. Solid line is from a
convolution of Eq. (1) with a Gaussian size distribution of the nanoparticles
various magnetic materials, calculated using theories for do- and using the 250 K data as background.
mains in fine particles,7,9 and using bulk properties tabulated
in the literature.10 Notice that due to surface effects for nano-
particles compared to bulk material, the anisotropies are gen- wise, if the magnetic relaxation is due to the Néel mecha-
erally higher, while the saturation magnetizations tend to be nism the low frequency peak should not have disappeared at
lower.11 Thus, Fig. 1 provides only an approximate guide for 250 K. A fit to Eqs. (1) and (2) with a Gaussian size
the needed particle size. Nevertheless, independent of the distribution7 suggests a ±12% distribution of the diameter of
magnetic material, the suitable magnetic particle size is typi- the nanoparticles involved in the Brownian relaxation. Note
cally of the order of a few tens of nanometers. that the high frequency responses at the two different tem-
We started with the magnetic characterization of avidin- peratures are similar regardless of the presence of the low
coated magnetite 共Fe3O4兲 nanoparticles in aqueous frequency peak. This is probably due to the existence of
some superparamagnetic particles since the median diameter
solution.12 The particle concentration in the fluid is
of the magnetic core 共⬃10 nm兲 is close to the crossover
⬃6 mg/ ml (~2⫻ 1015 particles/ml). Transmission electron
microscope images of dried samples show a magnetite core regime between Brownian and Néel relaxation (see Fig. 1).14
of ⬃10 nm diameter covered by an avidin coated shell that is In order to demonstrate the sensing scheme, we used
20– 30 nm thick. For the measurements, the nanoparticle commercially available biotinylated S protein.15 The specific
interaction of biotin and avidin protein has been well char-
sample was diluted with phosphate buffer saline (PBS, pH
acterized in the literature with a large affinity constant on the
= 7.0). The ac magnetic susceptibilities of a 100 ␮l aliquot of
order of a femtomole.16 As shown in Fig. 3, upon adding
the solution were measured in a physical property measure-
biotinylated S protein 共6.3 ␮M兲 to the avidin coated magne-
ment system (PPMS).13 An ac amplitude of 10 Oe was ap-
plied for all the measurements, while the frequency was var- tite nanoparticles, the peak frequency decreases from 210 to
ied between 10 Hz and 10 kHz. 120 Hz. Since biotinylated S protein does not exhibit any
A room temperature hysteresis curve of avidin-coated magnetic properties, this frequency shift has to be induced
magnetite nanoparticles shows paramagnetic behavior. As by the interaction of biotinylated S protein with avidin-
shown below, this paramagnetic behavior is due to single
domain nanoparticles rotating freely in the liquid to be
aligned with the external magnetic field. The hysteresis curve
can be fitted to the classical model of paramagnetism
(Langevin function) in order to estimate the magnetic mo-
ment of each nanoparticle. From this fit the magnetic mo-
ment for each nanoparticle is 2.0⫻ 104 ␮B, which corre-
sponds well to the estimated magnetic moment of 2.8
⫻ 104 ␮B for a spherical magnetite nanoparticle with 10 nm
diameter derived from literature data for the magnetization
of bulk magnetite.
Figure 2 shows the imaginary part of the magnetic sus-
ceptibility as a function of frequency of the external applied
magnetic field. At 300 K there is a peak in the ac suscepti-
bility at 210 Hz. When the PBS buffer solution was cooled to
250 K, which is below its freezing point, the frequency peak
disappeared (see Fig. 2). Since the freezing of the liquid
FIG. 3. Imaginary part of the ac magnetic susceptibility of an avidin-coated
immobilizes the nanoparticles at 250 K, this implies that the magnetic particle before (solid) and after (open) binding to S protein. Dif-
low frequency peak at room temperature is due to rotational ferent concentrations of S protein (open circles and squares) result in similar
diffusive Brownian relaxation of the magnetization. Other- frequency shifts.
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Appl. Phys. Lett., Vol. 85, No. 14, 4 October 2004
FIG. 4. Imaginary part of the ac magnetic susceptibility of an avidin-coated
magnetic particle before (solid) and after (open) binding to biotinylated T7
bacteriophage.
coated magnetic nanoparticles. Such an interaction will con-
sequently lead to the increase of the nanoparticles’ hydrody-
namic radius. From Eqs. (1) and (2), the peak frequency of
the ac magnetic susceptibility is inversely proportional to the
particle volume, i.e., ␻ p ⬀ 1 / r3. Assuming the diameter of the
avidin-coated magnetite particles is about 50 nm, as deter-
mined from the TEM measurements, this relation implies
that the particle diameter after S protein binding is about
60 nm. This increase corresponds well to the size of the S
protein (4 nm, estimated from the crystal structure of its
“parent” protein–ribonuclease A). When ten times more S
proteins were added to the sample, the peak frequency was
further reduced. However, the change was much smaller than
that for the initial addition of S protein since most of the
nanoparticle surface was covered by the 1 : 1 mixture with S
protein. This result demonstrates the feasibility of using the
frequency peak of the ac susceptibility to monitor the attach-
ment of the targeted molecules to magnetic nanoparticles,
and thus provides a biosensor scheme based on the Brownian
relaxation of magnetic nanoparticles in a liquid.
To further test our approach, we pretreated the biotiny-
lated S protein with S peptide displayed T7 bacteriophage
particles (415 copies of S peptide per phage) so that the
biotinylated S protein will be anchored to the surface of the
T7 bacteriophage due to the specific interaction between the
S peptide and S protein. Subsequently such biotinylated T7
bacteriophage particles were added to the magnetic nanopar-
ticle solution. The resulting ac susceptibility measurements
are shown in Fig. 4. Interestingly, the addition of biotinylated
T7 bacteriophage suppresses the peak in the imaginary part
of the ac susceptibility, similar to the effect of freezing of the
liquid (see Fig. 2). Thus this indicates that the magnetic
nanoparticles are immobilized upon binding to the T7 bacte-
riophage. 415 copies of biotin molecules per phage particle
can effectively crosslink the avidin-coated magnetite nano-
particle and thus causes their aggregation, which actually
immobilizes the particles. Thus the rotational motion of the
particles in the aggregate is blocked causing the disappear-
ance of the frequency peak by Brownian relaxation. It should
be pointed out that Weitschies et al.17 reported the observa-
tion of Néel relaxation via SQUID measurements due to the
immobilization of magnetic nanoparticles by biological bind-
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Chung et al. 2973
ing activity. The difference between this and the present
work is that the Brownian relaxation was outside of the
SQUID measurement window where the Néel relaxation
measurement was possible.
The main advantage of the sensing scheme presented
here is that the modification of Brownian relaxation offers an
opportunity to distinguish between various possible targets.
This is shown by the changes in the frequency response upon
binding to the S protein (see Fig. 3) and biotinylated T7
bacteriophage (see Fig. 4). In the first case, the binding re-
action results in a slower Brownian relaxation due to the
increased hydrodynamic radius, while in the second case the
crosslinking of the magnetic nanoparticles with the T7 bac-
teriophage gives rise to a suppression of the Brownian relax-
ation altogether. Furthermore, since the original magnetic
nanoparticles give rise to a characteristic frequency peak,
even in the absence of a suitable target, the sensing scheme
demonstrated here has therefore an inherent check for
integrity.
In conclusion, we have experimentally demonstrated the
detection of biomolecules using magnetic nanoparticles sus-
pended in a liquid. Our sensor scheme is an experimental
realization of the theoretically proposed approach of Con-
nolly and St. Pierre.7 The sensing of the target molecules is
based on the change of hydrodynamic radius due to the bind-
ing reaction with the magnetic nanoparticles. This increase in
the hydrodynamic radius leads to a characteristic decrease in
the peak frequency of the imaginary part of the ac magnetic
susceptibility. Therefore the sensing scheme has the advan-
tage of permitting one to distinguish between several poten-
tial targets. Future work will focus on increasing the sensi-
tivity and how well this sensing scheme can be applied to
heterogeneous mixtures of different targets.
The authors acknowledge stimulating discussions with J.
Meersschaut. This work was supported by the U. S. Depart-
ment of Energy, Basic Energy Sciences, under Contract No.
W-31-109-ENG-38 and DARPA, under Contract No.
8C67400.
1
For a recent review, see, Q. A. Pankhurst, J. Connolly, S. K. Jones, and J.
Dobson, J. Phys. D 36, R167 (2003).
2
M. M. Miller, G. A. Prinz, S.-F. Cheng, and S. Bounnak, Appl. Phys. Lett.
81, 2211 (2002).
3
P.-A. Besse, G. Boero, M. Demierre, V. Pott, and R. Popovic, Appl. Phys.
Lett. 80, 4199 (2002).
4
H. A. Ferreira, D. L. Graham, P. P. Freitas, and J. M. S. Cabral, J. Appl.
Phys. 93, 7281 (2003).
5
G. Li, V. Joshi, R. L. White, S. X. Wang, J. T. Kemp, C. Webb, R. W.
Davis, and S. Sun, J. Appl. Phys. 93, 7557 (2003).
6
A. Haller, S. Hartwig, H. Matz, J. Lange, T. Rheinländer, R. Kötitz, W.
Weitschies, and L. Trahms, Supercond. Sci. Technol. 12, 956 (1999).
7
J. Connolly and T. G. St. Pierre, J. Magn. Magn. Mater. 225, 156 (2001).
8
M. I. Shliomis, Sov. Phys. Usp. 17, 153 (1974).
9
C. Kittel, Rev. Mod. Phys. 21, 541 (1949).
10
Landolt-Börnstein (Springer, Berlin, 1994).
11
F. Bødker, S. Mørup, and S. Linderoth, Phys. Rev. Lett. 72, 282 (1994).
12
Liquids Research, Ltd., UK.
13
Quantum Design, San Diego, CA.
14
J. Zhang, C. Boyd, and W. Luo, Phys. Rev. Lett. 77, 390 (1996); P. C.
Fannin, B. K. P. Scaife, and S. W. Charles, J. Magn. Magn. Mater. 72, 95
(1988).
15
Novagen, Madison, WI.
16
N. M. Green, Adv. Protein Chem. 29, 85 (1975).
17
W. Weitschies, R. Kötitz, T. Bunte, and L. Trahms, Pharm. Pharmacol.
Lett. 7, 5 (1997).