Review TRENDS in Biotechnology
Biosensor developments: application
to prostate-specific antigen detection
Declan A. Healy1, Conor J. Hayes1, Paul Leonard1, Louise McKenna2 and
Richard O’Kennedy1
1
School of Biotechnology and Biomedical Diagnostics Institute, National Centre for Sensor Research, Dublin City University,
Dublin 9, Ireland
2
Forensic Science Laboratory, Garda Headquarters, Phoenix Park, Dublin 8, Ireland
Prostate-specific antigen (PSA) is the best serum
marker currently available for the detection of prostate
cancer and is the forensic marker of choice for deter-
mining the presence of azoospermic semen in some
sexual assault cases. Most current assays for PSA
detection are processed on large analyzers at dedi-
cated testing sites, which require that samples be sent
away for testing. This leads to delays in patient man-
agement and increased administration costs. The
recent emphasis placed on the need for point-of-care
patient management has led to the development of
novel biosensor detection strategies that are suitable
for the miniaturization of assays for various targets
including PSA. This review highlights the current and
novel analytical technologies used for PSA detection,
which will benefit clinicians, patients and forensic
workers in the future.
Introduction
Prostate cancer (PCa) accounts for 10% of all deaths
from cancer [1–4], and the indicator most widely used to
detect it is serum prostate-specific antigen (PSA) levels:
PSA is a serine protease that is produced by the prostate
epithelium to maintain liquefaction of seminal fluid [5].
The importance of PSA as an oncological marker is
partly because of the lack of real alternative markers
of PCa. Most PSA testing takes place at dedicated cen-
tralized laboratories using large, automated analyzers,
requiring sample transportation, increased waiting
times and increased administration and medical costs.
The availability of near-patient or point-of-care testing
(POCT) could help to reduce the number of clinic visits,
decrease costs to the patient and the healthcare system,
increase patient satisfaction and improve clinical out-
come. Recent advances in biosensor development, using
nanoparticles and nanostructures as integral com-
ponents, have brought POCT for PSA closer to reality.
This review highlights some important biosensor formats
that have recently been applied to PSA detection and
emphasizes the advantages or disadvantages of each
biosensor type with respect to its suitability for use in
the POCT devices of the future.
Corresponding author: O’Kennedy, R. (Richard.OKennedy@dcu.ie).
Available online 24 January 2007.
www.sciencedirect.com 0167-7799/$ – see front matter ß 2007 Elsevier Ltd. All rights reserved. doi:10.1016/j.tibtech.2007.01.004
Vol.25 No.
Prostate cancer detection using serum PSA
measurement
Trace levels of PSA are naturally found in the serum;
however, PCa tumor growth usually leads to the release
of high concentrations of PSA into the circulatory system
[6]. A PSA measurement above a cut-off value of 4.0 ng/ml
(and more recently 2.5 ng/ml) is generally regarded as
positive and might indicate the need for a biopsy. PSA
testing is also used to monitor the response of PCa patients
to ablative therapy such as radical prostatectomy. In this
situation PSA should be undetectable, and a subsequent
detectable level of PSA is a sign of disease recurrence. For
this reason, ultrasensitive assays that are capable of
detecting concentrations of serum PSA in the low pg/ml
region have been sought to facilitate earlier detection of
recurrence.
Despite its wide use, PSA is not a cancer-specific
marker, and other non-cancerous diseases of the prostate,
such as benign prostatic hyperplasia (BPH), can also lead
to increased release of PSA into the circulation. The devel-
opment of assays that are capable of distinguishing
between different isoforms of PSA [1,7], such as the ratio
of free PSA to complexed PSA, has helped to improve the
distinction between PCa and BPH, particularly in the so-
called ‘diagnostic grey zone’ of 4–10 ng/ml PSA [6–10].
PSA detection in forensic samples
In addition to its use in the diagnosis of PCa, the detection of
PSA has also become the method of choice for the forensic
determination of the presence of semen, in the absence of
sperm, in sexual assault cases. However, the detection of
PSA in forensic samples necessitates different assay
requirements than its detection in clinical samples. First,
it is not the determination of absolute PSA levels that is
important in forensic science, rather the ability to detect
PSA in what are invariably ‘dirty’ samples: contamination
with other body fluids or dirt, scarcity of the sample, the need
for extraction from different types of fabric – leading to low
extraction efficiencies – and the decomposition of samples in
cadavers or following laundering of fabrics are all common
challenges to PSA detection in forensic samples [11]. Second,
owing to low concentrations of PSA being naturally present
in the tissues and body fluids of some females, PSA assays
for forensic use require only limited sensitivity to avoid the
danger of false-positive results.
126 Review TRENDS in Biotechnology Vol.25 No.3

Table 1. Examples of some commercially available tests commonly used for PSA determinations in serum
Manufacturer Assay PSA isoform detected Lowest detection Expected range Sample Assay time Sample
limit (ng/ml) volume (mins) throughput
(ng/ml) required (ml) per hour
Abbott Diagnostics Architect Free PSA <0.008 0–30 NS 29 200
Total PSA <0.008 0–100 NS 29 200
AxSYM Free PSA <0.02 0–10 NS 19 62
Total PSA <0.04 0–50 NS 19 62
IMx Total PSA <0.04 0–50 NS 45 32 a
Diagnostic Products Immulite Total PSA 0.04 0–150 10 30 200
Corporation Free PSA 0.02 0–25 10 30 200
Third-generation PSA b 0.003 0–20 50 60 NS
Roche Elecsys Total PSA 0.002 0–100 5–50 18 85
Free PSA 0.01 0–50 5–50 18 85
Beckman Coulter Access-Hybritech Total PSA <0.008 0–150 25 20 NS
Free PSA <0.005 0–20 25 20 NS
Bayer Diagnostics ADVIA PSA-ACT 0.03 0–100 NS NS 240
a
Actual sample throughput is 24 samples in 45 min. Abbreviation: NS, not specified on product package insert. bThird-generation PSA assays are sensitive to 0.005 ng/ml,
compared with first-generation (0.5 ng/ml) and second-generation (0.05 ng/ml) assays.

Currently available PSA immunoassays
Currently, most PSA testing takes place at dedicated,
centralized laboratories on large, automated high-through-
put systems, and numerous analyzer-run PSA assays are
currently available in the marketplace (Table 1). The
advantages of such systems include low detection limits
(in the region of 0.05–0.005 ng/ml), proven reliability and
high-throughput of samples. However, an important dis-
advantage associated with the fact that these large sys-
tems are only found in dedicated laboratories (Table 2) is
the requirement for sample transportation to the testing
site: this results in delays in processing and reporting the
results back to the clinician. Thus, the time between the
decision to test and the therapeutic action taken on the test
result, termed the therapeutic turnaround time (TAT) [12],
is often of the order of several weeks. The ideal situation,
where results are returned within a matter of minutes,
could be facilitated through the introduction of portable
POCT devices. Patient experience would be revolutionized
through a reduction in the number of clinic visits,
decreased costs, increased patient satisfaction and
improved clinical outcome [13], similar to that seen for
other non-emergency diseases such as diabetes. Further-
more, the portability of such devices would have important
implications for the possibility of high-quality testing of
forensic samples at crime scenes.
In an effort to provide a POCT alternative to centralized
PSA testing, immunochromatographic membrane tests,
also known as immunostrip tests, have been developed.
The major advantage of immunostrip devices is their
low cost, robust nature and ease of use; however, immuno-
strip devices have not proved to be a real alternative to
centralized laboratory testing for clinical PSA samples
because of their general lack of sensitivity and, at best,
semi-quantitative nature (Table 3). Despite this, PSA
immunostrip devices have been applied to PSA detection
in forensic semen samples, particularly in rape cases. The
relatively low sensitivities of immunostrip devices help to
avoid the false-positive results caused by the low levels of
PSA that can be present in many female tissues and fluids.
Although more suited for use with forensic samples than
centralized immunoassay tests, membrane tests still suffer
from several problems related to specificity, false positives,
interference from matrix components and the instability of
PSA [14–16].
Biosensor development: a trend towards point-of-care
PSA measurement
An ongoing revolution in the area of medical diagnostics is
the development of biosensors that have the potential for
use in POCT devices. These would provide rapid and
reliable quantitative results ‘anytime, anywhere’, for
example, clinics and hospital emergency departments. In
order for POCT to become a reality, current biological
testing formats must be reduced to the size of handheld
devices that require only small amounts of sample and
reagents, while maintaining a multi-analyte, high-
throughput, specific and sensitive assay. Recent advances
in biosensor development, using nanoparticles and nanos-
tructures as integral components, have enabled the devel-
opment of analyte-detection methods in the nanoscale
(Figure 1). These have obvious advantages for use in POCT
settings compared with traditional measurement tech-
niques. A biosensor is an analytical device that brings

Table 2. Limitations of current PSA testing technologies

Features of current PSA immunoassays Limitations
Run on large, high-throughput analyzers  High cost of analyzer and associated instrumentation
 Restricted to dedicated laboratories
 Up to 1 ml of sample required per test
Testing at dedicated laboratories  Requirement for sample transportation at high cost
 Requirement for storage until testing
 Several weeks from testing to results
Complicated technology  Requirement for highly trained technical personnel
Thousands of samples from different sources analyzed per day  High administration costs
 Potential for sample mix-ups
Each PSA isoform measured using different reagents  Many sets of reagents required, leading to high cost

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 Review TRENDS in Biotechnology Vol.25 No.3 127

Table 3. Performance characteristics of some commercially available immunostrip PSA tests

PSA strip test Manufacturer Sample type Compared with PSA Sensitivity Specificity Refs
cut-off (ng/ml) (%) (%)
Biosign PSA test Princeton BioMeditech Corp, Serum Tandem-E 4.0 63 92 [43]
USA Abbott IMX 4.0 68 95
Chembio Chembio, USA Serum DPC Immulite 4.0 67 87 [44]
Medpro Medpro, UK Serum DPC Immulite 4.0 87 88 [44]
Coulter Hybritech 3.1 64 75 [45]
PSA semiquant test Seratec Diagnostica, Serum DPC Immulite 4.0 80 97 [44]
Germany
OneStep PSA test Syntron, Serum DPC Immulite 4.0 93 93 [44]
USA
Onco-screen FF Diagnostics, Serum Tandem-E 4.0 76 93 [46]
PSA test Germany
PSA strip test Cardimac GmBh, Whole blood Abbott IMX 4.0 91 84 [47]
Germany

together an immobilized biological sensing material and a
transducer, to produce a quantifiable signal that is pro-
portional to the concentration of the analyte. Biosensors
comprise three distinct components: (i) a biorecognition
element, usually an antibody, to recognize and bind the
analyte or analytes of interest; (ii) a transducing element
that converts the interactions of biomolecules (e.g. anti-
body and antigen) into a quantifiable signal; and (iii) a
readout system. The development of biosensors for the
measurement of PSA has almost exclusively used the
binding of PSA by high-affinity intact antibodies to form
the recognition element of the devices. Descriptions of all
the newly developed biosensor formats is outside the scope
of this short review; therefore, the following sections high-
light some of the more important signal transduction
developments that have been applied to PSA biosensor
development and the trend towards the use of label-free
modalities (Table 4). Particular emphasis is given to the
advantages or disadvantages of each signal transduction
method with respect to multiplexing potential and its
suitability for incorporation into POCT devices. The suit-
ability of biosensors for the multiplexed analyses of several

Figure 1. Schematic representations of some biosensor formats applied to PSA detection. (a) Immuno-PCR: after PSA capture by anti-PSA antibodies on the biosensor
surface, a DNA-labeled detection antibody binds. After washing steps, DNA is amplified by real-time PCR, where during the exponential phase of the PCR the amount of
product formed reflects the amount of PSA in the sample. Adapted from [24]. (b) Biobarcode assay: PSA binds to anti-PSA antibodies coated onto magnetic microparticles.
Nanoparticle probes coated with anti-PSA antibody and barcode-DNA strands are used to bind immobilized PSA. Following washing, barcode DNA is dehybridized from the
nanoparticle probes and quantified. Adapted from [26]. (c) SERS: binding of PSA to anti-PSA antibodies functionalized with Raman reporter molecules (RRMs) and
immobilized onto gold microparticles leads to altered spectroscopic output of the RRMs, which can be used to quantify the concentration of PSA. Adapted from [21].
(d) Electrical detection of micro-cantilever bending: piezoresistive microcantilevers, functionalized with anti-PSA antibodies, bend owing to surface stress caused by PSA
capture. Micro-cantilever bending is measured by electrical detection. Adapted from [40].

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128 Review TRENDS in Biotechnology Vol.25 No.3

Table 4. Examples of biosensor formats applied to PSA detection

Signal transduction Assay format and/or nanostructure used Lowest detection Sample Assay time Refs
limit volume (mins)
required
(ml)
Time-resolved Europium (III) chelate-dyed polystyrene nanoparticles 0.21 ng/ml 20 240 [48]
fluorescence Silica-coated terbium (III) fluorescent nanoparticles 7.0 pg/ml 45 >180 [18]
Terbium complex-doped zirconia nanoparticles 0.4 ng/ml 45 >180 [20]
Europium(III) nanoparticle labels and streptavidin–biotin technology 0.83 pg/ml 5 >150 [19]
SERS Gold nanoparticles 1.0 pg/ml 40 550 [21]
Real-time Immuno-PCR Sandwich assay with DNA label on detection antibody 0.2 pg/ml a 5.5 >150 [24]
Immuno-RCA Sandwich assay with DNA label on detection antibody 0.1 pg/ml 10 180 [25]
Biobarcode Gold nanoparticles 1.0 fg/ml b 10 80 [26]
1.0–10.0 fg/ml 250 210 [27]
Enzyme and/or Lateral flow immunostrip containing an electrochemical transducer 3.0 ng/ml 11 10–30 [29]
impedance
Enzyme and/or Sandwich immunoassay on three-electrode system 0.25 ng/ml 10 60 [31]
amperometric
Surface plasmon Commercial SPR biochip with signal enhancement using a sandwich- 18.1 ng/ml 100 14 [35]
resonance assay format 1.0 ng/ml 130 6.5 [34]
0.15 ng/ml NS NS [49]
Surface plasmon Sandwich-immunoassay format on commercial SPR biochip 3 pg/ml 500 45 [50]
fluorescence
spectroscopy
Electrical Resonance frequency shift in a nanomechanical microcantilever 10 pg/ml NS NS [36]
100 ng/ml 20 60 [37]
Surface stress bending of piezoresistive self-sensing microcantilevers 10 ng/ml NS 10 [38]
Conductance change in silicon nanowire sensor chip 100 fg/ml NS 30 [40]
a
Sensitivity reported by authors as 4.8  105 PSA molecules, converted to pg/ml for consistency; bsensitivity reported as 30 aM, converted to fg/ml for consistency.
Abbreviation: NS, not specified by authors.

markers simultaneously will be particularly important for
distinguishing PCa from benign diseases, such as BPH,
and for facilitating better detection of PSA in forensic
samples, through the incorporation of antibodies against
epitopes of PSA that are more easily identified in contami-
nated samples.
Label-based signal transduction
Fluorescent labels
Although fluorescence-dependent signal generation has
disadvantages with regard to its use in multiplexed assays,
fluorescence continues to be a major transduction modality
in biosensors for the detection of PSA using optical readout
systems. In particular, fluorescent nanoparticles, such as
quantum dots and luminophore-doped nanoparticles, have
provided an attractive field of research for biosensor devel-
opment because they have high photostability and narrow
emission peaks, enabling greater multiplexing potential.
With respect to PSA assay development, europium(III)-
and terbium (III)-doped fluorescent polystyrene and zirco-
nia nanoparticles have been used for the detection of PSA
to a concentration <1 pg/ml [17–20].
Surface-enhanced Raman spectroscopy
Surface-enhanced Raman scattering (SERS) has been used
in biosensor development as an alternative to fluorescence
labeling. Raman dyes are used to label antibodies that are
attached to gold nanoparticle probes; the light scattered
from the Raman reporter molecule (RRM) provides infor-
mation about the vibrational quantum states of a molecule.
SERS was used in a sandwich-immunoassay format for the
detection of PSA to a concentration of 1 pg/ml [21]. SERS
was also used for the detection of multiple analytes [22]:
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this is possible because Raman bands are sufficiently
narrow to reduce the likelihood of spectral overlap when
multiple labels are used.
DNA labels as surrogate markers: from immuno-PCR to
biobarcode assays
DNA tags have been used as signal amplification and
transducing mechanisms in biosensor development for
the past 15 years. They have great potential for use in
multi-analyte detection systems because different specific
sequences can be associated with each individual target
analyte of interest. The first DNA label-based technique
was known as immuno-PCR [23]; and this used a secondary
antibody labeled with a DNA molecule that could be
amplified by PCR as a signal amplification method. More
recently, real-time PCR, using fluorescence measurements
to follow the DNA amplification, has been used in biosen-
sors to detect 4.8  105 PSA molecules in a 5 ml sample –
equivalent to a sensitivity of 0.2 pg/ml [24]. Although
suitable for multiplex protein quantification, real-time
immuno-PCR-based biosensors are currently hindered
by long assay times (3–4 h) and the need for the incorp-
oration of multiple components and thermal cycling.
Rolling-circle amplification (RCA) was used as a reporter
system for the sensitive detection of PSA, in an assay
termed immuno–RCA. An oligonucleotide primer is
attached to a reporter antibody that is specific for the target
analyte and is then amplified to generate an extended DNA
molecule containing many copies of the circular DNA
sequence, which remains attached to the reporter antibody
for detection by fluorescent imaging. PSA has been detected
at 0.1 pg/ml in a microspot array assay using this technique
[25].
 Review TRENDS in Biotechnology
A highly sensitive, DNA-labeled, nanoparticle-based
assay system that does not require signal amplification
has recently been developed [26]. This ‘biobarcode’ tech-
nology relies on the use of magnetic beads that are func-
tionalized with antibodies – to capture the target of
interest – and nanoparticle probes with both a DNA oli-
gonucleotide sequence (the barcode) that is unique for each
of the target proteins and a specific antibody that binds, in
a sandwich-assay format, to the target protein, already
captured by the antibody on the magnetic beads. Following
magnetic separation of the complex, the target protein
concentration is determined by measurement of the oligo-
nucleotide (barcode) levels, which are directly related to
the levels of the target protein. PSA was detected at 1 fg/ml
(30 aM) concentration in a 10 mL sample, in a PCR-less
biobarcode assay [26]. A recent modification to the barcode
assay for PSA detection has been reported [27]; here, a
brief exposure to silver stain increased the size and light-
scattering properties of nanoparticle tracers, enabling PSA
to be detected at 1–10 fg/ml with low resolution optics and
without the need for laser excitation. The high sensitivity
and huge multiplexing attributes of biobarcode assays are
particularly suitable for use in POCT devices.
Electrochemical signal transduction
Electrochemical devices have received much attention in
the field of biosensor development because they provide a
simple, inexpensive and accurate platform for the
measurement of the target analyte [28]. Electrochemical
biosensors determine the level of the analyte by detecting
the changes in either potential, current, capacitance, con-
ductance or impedance that are caused by a specific bior-
ecognition reaction.
The development of a rapid, single-use, lateral flow,
nitrocellulose immunostrip test device, which is based
on capacitance measurement, has recently been reported
[29]. This device incorporates an electrochemical transdu-
cer coated with a pH-sensitive polymer layer to enable
measurement of PSA to a sensitivity of 10 ng/ml: binding of
PSA to an immobilized anti-PSA antibody and an anti-PSA
antibody–urease conjugate catalyzes the hydrolysis of ur-
ea, thereby generating ammonia, leading to an increase in
the pH of the reaction solution. This leads to the degra-
dation of the polymer layer coating the transducer, which
can be monitored as a change in the capacitance of the
system. In addition to being robust and inexpensive to
make, this system has the advantage of requiring just one
manual step: the addition of 11 ml of sample. The use of
impedance- and capacitance-based immunosensors and
immunostrip devices in future multiplexing POCT devices
might be particularly suited to forensic applications,
although significant improvements in their sensitivities
will be required before they are used on clinical samples.
Amperometric biosensors are based on the measure-
ment of the current associated with an electrochemically
active species that is produced by, in general, an enzyme
label. Amperometric detection of enzyme labels has
the advantage of providing higher sensitivity than impe-
dance and capacitance methods and uses accurate sensors
that are simple and inexpensive to produce. An example of
an amperometric immunosensor for PSA detection was
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Vol.25 No.3 129
presented as far back as 1995 [30]. More recently, there
has been a trend towards the use of screen-printed electro-
des owing to their simple and low-cost fabrication and ease
of use. For example, amperometric detection of PSA at
0.25 ng/ml using a three-electrode system has been
reported [31].
Although amperometric biosensors currently suffer
from the necessity for washing and separation of the free
from the bound label, simplification of these biosensors
holds great promise for their incorporation into POCT
devices of the future. Recent advances, such as the devel-
opment of a reagentless amperometric immunosensor
based on direct electrochemistry enzyme labels and the
testing of microfluidic devices using amperometric detec-
tion strategies on microchip platforms, might be of great
importance to POCT devices of the future. Furthermore,
the suitability of amperometric biosensors to multiplex
analyte detection has already been demonstrated, where
electrode arrays with multiple, individually addressable
electrochemical transducers have been fabricated [32].
Label-free transduction
Biosensors that use label-free transduction methods do not
require a reporter molecule to signal the presence of target
analyte on the sensor surface, thereby enabling the use of
unmodified samples, with the possibility of ‘real-time’
measurement. Examples of some label-free methods used
in PSA biosensors are described below.
Surface plasmon resonance
Many of the biosensors currently available on the market-
place use the optical properties of lasers to monitor the
interactions of biomolecules. Surface plasmon resonance
(SPR) is an optical–electrical technology that involves
the interaction of light with the electrons of a metal.
The technique enables rapid, label-free monitoring of bio-
molecular reactions in real-time, and requires only a small
amount of sample. SPR measures changes in the refractive
index and thus in the resonance angle at which polarized
light is reflected from a surface, which is, in turn, related to
a change in layer thickness or mass upon antigen binding
to an antibody-coated gold layer [33].
SPR-based detection of different PSA isoforms to 1 ng/
ml using a commercial SPR biosensor (Biacore AB; http://
www.biacore.com/) has been reported by several groups
[34,35]. However, the Biacore system is expensive to pur-
chase and unsuitable for applications that require port-
ability owing to its size. Recent advances in the
miniaturization of SPR technology have made possible
the development of portable systems that strengthen
the probability of SPR biosensor incorporation into future
POCT devices. The Spreeta 2000 SPR biosensor elements,
manufactured by Texas Instruments (http://www.ti.com/),
enables the simultaneous label-free analysis of multiple
analytes in a portable format. Furthermore, biosensors
that use the SPR of noble metal nanoparticles and nanos-
tructures, known as nanoSPR, for the optical transduction
of biorecognition events have been developed. In nanoSPR,
visible light induces the collective oscillations of the sur-
face electrons and is responsible for the intense colors
emitted by colloidal solutions of noble metals, including
130 Review TRENDS in Biotechnology Vol.25 No.3
gold and silver [36]. In particular, nanoSPR can be applied
to a chip format, in which nanoparticles with different
optical signatures and coated with different antibody spe-
cificities [37] are immobilized in an array format for high-
throughput screening of biomolecular binding interactions
in a real-time, label-free manner.
Electrical and electromechanical signal transduction
The advent of nanostructures that act as signal
transducers in a label-free manner undoubtedly show
the most promise for use in POCT devices of the future.
Nanostructure-based assays will prove to be both robust
and inexpensive to produce owing to the elimination of the
need for sample preparation and labeling steps.
Micro-cantilever biosensors do not require a reporter
molecule to signal the presence of the target analyte on the
sensor surface, thereby enabling the use of unmodified
samples, with the possibility of real-time measurement.
Microcantilevers are particularly suited to incorporation in
POCT devices owing to their minute size (100 mm2), and
lend themselves to the possibility of microcantilever
arrays, enabling multiplex analysis of numerous markers.
Microcantilever biosensors can be operated in one of two
modes: resonance response variation (microbalance
method) and cantilever bending (surface-stress method).
Using the microbalance method, Kim et al. have reported
that electrical measurement of the resonant frequency
change of an anti-PSA-antibody-coated microcantilever
upon PSA binding enables detection of PSA in solution
at concentrations as low as 10 pg/ml [38,39]. Unfortu-
nately, the microbalance method suffers from an undesir-
able resonant frequency change due to variations in the
viscosity of the medium and cantilever damping in a liquid
environment [40]. Therefore, Kim’s group have sub-
sequently used electrical detection of cantilever bending
due to surface-stress changes, to measure PSA-antibody
binding on the surface of micro-fabricated, self-sensing
microcantilevers, with a reported sensitivity of 10 ng/ml
[40]. Microcantilever technology is particularly suited to
miniaturized biosensor development because it is a rapid,
sensitive, specific, cost-effective and portable technique
that does not require labeling or external excitation. In
addition, it has been demonstrated that microcantilevers
can be used in an array format for the sensitive and
simultaneous measurement of multiple unlabeled analytes
[41]. The incorporation of cantilever technology into micro-
fluidic channels will reduce the amount of sample required.
Silicon nanowire field-effect sensors coated with anti-
body have been used for the real-time, highly sensitive
detection of PSA [42]. Silicon nanowires act as signal
transducers through a change in conductance upon binding
of a target analyte to the antibody receptor on the nanowire
surface. They have been used by Zheng et al. to detect PSA
to a concentration of 0.9 pg/ml [42]. This group demon-
strated the multiplexing ability of their nanowire sensor
chip by simultaneously detecting both free PSA (fPSA) and
bound PSA (PSA–ACT bound) at pg/ml concentrations.
Furthermore, the authors report the real-time multiplexed
detection of the cancer markers fPSA, carcinoembryonic
antigen (CEA) and mucin-1 at concentrations in the range
of 50–100 fg/ml – attributes that are likely to be important
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in future POCT devices for the screening of numerous
diseases using a single blood sample.
Future perspectives and conclusions
Recent developments in biosensor technology have enabled
the transition from centralized to near-patient PSA testing
to emerge from the realms of possibility and become closer
to reality. In future, results will be available within min-
utes, thereby revolutionizing the patient experience
through reduced hospital or clinic visits, decreased costs
and improved clinical outcomes. The use of nanoparticle-
based assays for PSA, comprising indirect protein ampli-
fication technologies such as immuno-PCR and biobarcode
assays, electrochemical assays based upon molecular
probes, and assays using reporter molecules such as RRMs,
offer higher sensitivities than traditional ELISAs on a
miniaturized scale. The trend towards the use of label-free
biosensor formats, including SPR, microcantilever and
nanowire-based assays, for PSA detection, enable the
use of unmodified samples, with the possibility of highly
sensitive real-time measurement. In addition to the need
for multi-analyte-measuring capabilities, the success of a
particular biosensor format in future POCT devices will
depend on the ability to incorporate the format into devices
that comprise simple and efficient sample handling and
readout systems. Such laboratory-on-a-chip, or micro-total
analysis systems (mTAS), will incorporate the functions of
a full-scale laboratory in a portable device that will provide
rapid and accurate sample analysis of minute sample
volumes in every environment.
Acknowledgements
We thank the following for financial support: Enterprise Ireland (DAH),
Cancer Research Ireland (CJH) and Science Foundation Ireland (PL).
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