2013年SPR核酸检测技术综述：生物分析与分子互动研究进展
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本文是一篇2013年发表在《分析化学 Acta》上的综述文章，标题为《表面等离子共振检测核酸：一次回顾》（Surface plasmon resonance sensing of nucleic acids: A review）。作者Hanaˇ Sípová和Jiˇ rí Homola来自捷克共和国科学院的光子学与电子学研究所。该研究对当时关于核酸（NA）表面等离子共振（SPR）传感器的最新进展进行了详尽的探讨。
文章首先强调了核酸表面等离子共振传感器的显著进步，这些传感器在生物分析学领域的应用越来越广泛。它们能够精确地检测和识别核酸分子，这对于基因组学、蛋白质组学以及疾病诊断等领域具有重要意义。通过 SPR 技术，研究人员可以实时监测 DNA、RNA 或寡核苷酸与目标分子之间的相互作用，从而深入了解分子间的交互作用机制。
作者详细梳理了此类传感器的设计原理，包括如何利用金属-介质界面处的表面等离子体振动，当与特定的核酸分子结合时，会导致 SPR 波长发生微小变化，这一现象被用来作为生物分子存在与否的信号。这种灵敏度和选择性使得SPR成为分析生物大分子的理想工具。
此外，文中还涵盖了多项实际应用案例，展示了SPR在生物标志物检测、病原体检测、药物筛选以及基因表达调控研究中的应用。例如，它在疾病早期诊断中的潜力，如癌症和感染性疾病，以及在纳米技术中作为生物传感器在纳米药物递送系统中的作用。
文章最后总结了当时的科研热点，指出尽管取得了显著的进步，但仍存在一些挑战，如提高传感器的稳定性、降低背景噪音和增强对复杂生物环境的耐受性。未来的研究方向可能集中在开发新型材料和设计策略，以优化SPR传感器的性能，并进一步拓展其在生物科学和医疗领域中的应用。
这篇综述为读者提供了一个全面的视角，深入剖析了SPR技术在核酸检测中的现状、应用潜力以及未来发展方向，对于理解该领域的最新研究成果和技术趋势具有很高的参考价值。
Please
cite
this
article
in
press
as:
H.
ˇ
Sípová,
J.
Homola,
Surface
plasmon
resonance
sensing
of
nucleic
acids:
A
review,
Anal.
Chim.
Acta
(2013),
http://dx.doi.org/10.1016/j.aca.2012.12.040
ARTICLE IN PRESS
G
Model
ACA-232306;
No.
of
Pages
15
H.
ˇ
Sípová,
J.
Homola
/
Analytica
Chimica
Acta
xxx (2013) xxx–
xxx 3
Fig.
1.
Concept
of
a
surface
plasmon
resonance
(SPR)
biosensor.
(A)
Excitation
of
PSPs
by
a
polychromatic
light
in
the
Kretschmann
geometry
of
the
ATR
method.
(B)
Spectrum
of
reflected
light
with
a
characteristic
SPR
dip
for
two
different
values
of
the
refractive
index
at
the
gold
layer.
(C)
SPR
sensor
surface
with
immobilized
probes
(left)
and
binding
of
the
target
molecules
to
the
probes
(right).
(D)
Temporal
evolution
of
the
refractive
index
caused
by
the
molecular
interaction.
coupler
via
the
attenuated
total
reflection
method
(ATR).
In
the
Kretschmann
geometry
of
the
ATR
method,
a
light
wave
passes
through
a
high
refractive
index
prism
and
excites
a
PSP
on
the
outer
boundary
of
the
metal
film
located
on
the
base
of
the
prism
(Fig.
1A).
The
excitation
of
the
PSP
gives
rise
to
a
drop
in
the
intensity
of
the
reflected
light.
A
change
in
the
refractive
index
of
the
dielectric
medium
in
the
evanescent
wave
of
the
PSP
produces
a
change
in
the
propagation
constant
of
the
PSP.
This
change
in
propagation
constant
alters
the
coupling
condition
between
the
light
wave
and
the
PSP,
which
can
be
observed
as
a
change
in
the
characteristics
of
the
optical
wave
(coupling
angle
of
incidence,
coupling
wavelength,
intensity
and
phase,
Fig.
1B).
In
the
last
two
decades,
a
large
variety
of
SPR
sensor
platforms
have
become
available
[21–24]
including
both
laboratory
systems
for
parallelized
monitoring
of
molecular
interactions
[25–29]
and
compact
SPR
sensors
for
bioanalytical
applications
in
the
field
[8,30–36].
A
typical
resolution
of
an
SPR
sensor
(i.e.
the
smallest
change
in
the
refractive
index
which
can
be
measured
by
the
sen-
sor)
is
of
the
order
of
10
−7
RIU
for
table-top
laboratory
systems
and
of
the
order
of
10
−6
RIU
for
portable
SPR
sensors
[10,20,37].
2.1.2.
SPR
affinity
biosensing
In
SPR
biosensors,
an
interacting
molecule
(probe)
is
immobi-
lized
on
to
the
surface
of
the
SPR
sensor
(Fig.
1C).
When
a
solution
containing
the
biological
counterpart
molecule
(target)
is
brought
into
contact
with
the
SPR
sensor,
the
target
molecules
in
solution
bind
to
the
immobilized
probes
via
affinity
interaction,
resulting
in
an
increase
in
the
refractive
index
at
the
SPR
sensor
surface
(Fig.
1D).
The
change
in
refractive
index
n
d
occurring
within
a
layer
of
thickness
h
can
be
expressed
as
n
d
=
dn
dc
vol
h
,
(1)
where
(dn/dc)
vol
is
the
increment
of
refractive
index
n
with
the
volumetric
concentration
of
analyte
c,
and
is
the
surface
con-
centration
of
the
bound
target
[38].
The
refractive
index
increment
(dn/dc)
vol
of
proteins
and
nucleic
acids
is
close
to
0.18
cm
3
g
−1
[39].
The
change
in
the
refractive
index
causes
a
change
in
the
coupling
of
incident
light
and
a
PSP.
By
tracking
the
characteristics
of
light
cou-
pled
to
a
PSP
in
real
time,
kinetic
details
of
the
interaction
between
the
target
molecules
in
the
solution
and
probes
immobilized
on
the
sensor
surface
can
be
determined.
In
addition,
the
refractive
index
change
caused
by
the
binding
of
target
molecules
may
be
used
to
identify
the
target
molecules
and
quantify
their
concentrations.
The
limit
of
detection
(LOD)
of
SPR
biosensors
is
typically
defined
as
a
concentration
of
analyte
producing
the
sensor
response
equal
to
three
standard
deviations
of
the
response
to
the
blank
sample.
In
the
state
of
the
art
SPR
biosensors,
the
analyte
concentra-
tion
corresponding
to
the
LOD
generates
the
surface
concentration
of
the
bound
target
of
about
0.3
pg
mm
−2
[40].
The
relationship
between
the
surface
concentration
of
the
bound
targets
and
the
volumetric
concentration
of
targets
is
given
by
a
set
of
diffusion
and
kinetic
equations
[40].
These
equations
suggest
that
a
state
of
the
art
SPR
biosensor
detecting
a
short
15-mer
target
NA
(MW
=
5
kDa)
via
a
complementary
probe
(k
a
=
10
4
M
−1
s
−1
,
k
d
=
10
−5
s
−1
[41])
is
able
to
detect
the
target
with
the
LOD
of
400
pM
[40].
2.1.3.
Investigating
NA
interactions
The
most
widely
used
model
of
NA
hybridization
on
solid
surface
assumes
that
the
interaction
between
a
single-stranded
nucleic
acid
(probe
p)
immobilized
on
the
surface
of
an
SPR
sensor
and
another
single-stranded
nucleic
acid
(target
t)
in
the
solution,
can
be
described
as:
p
+
t
k
a
←→
k
d
pt,
(2)
where
k
a
and
k
d
are
the
association
and
dissociation
rate
constants,
respectively.
The
surface
concentration
ˇ
of
p–t
complexes
can
be
expressed
as:
dˇ
dt
=
k
a
c
t
(ˇ
max
−
ˇ)
−
k
d
ˇ
(3)
where
c
t
is
the
concentration
of
the
target
in
the
proximity
of
the
sensor
surface,
ˇ
max
is
the
surface
concentration
of
the
probes,
and
t
is
time.
Eq.
(3)
describes
the
generally
accepted
model
of
1:1
inter-
actions
on
the
surface
of
an
SPR
biosensor.
It
describes
the
majority
of
the
NA
interactions
with
other
types
of
molecules,
such
as
inter-
calators,
minor
groove
binders
or
proteins.
Eq.
(3)
can
be
expanded
with
additional
terms
to
account
for
more
complex
molecular
inter-
actions
of
nucleic
acids,
such
as
intramolecular
interactions
(the
formation
of
secondary
structures),
the
formation
of
triple
helixes
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