Transcript Document

Advanced Biosensors Based on
Nano-scaled Arrays and Films of
Electro- and Biocatalysts
Arkady A. Karyakin
Chemistry faculty of M.V. Lomonosov Moscow State University, 119991, Moscow,
Russia
Electroanalysis:
• widest dynamic range
(down to a single molecule detection)
• less sensitive to matrix effects
(insensitive to color, turbidity etc.)
• the cheapest equipment
2
Scheme of biosensor action
Transducer
Biorecognition
element
Coupling of biochemical and
transducer reactions
Substrate recognition
Signal processing
3
Advanced biosensors:
•most efficient electrocatalyst
(transducer)
A.A.Karyakin, O.V.Gitelmacher, E.E.Karyakina. Analytical Chemistry 67 (1995) 2419
A.A.Karyakin, E.E.Karyakina, L.Gorton. Electrochem. Communications 1 (1999) 78.
A.A.Karyakin, E.E.Karyakina, L.Gorton. Analytical Chemistry 72 (2000) 1720
A.A.Karyakin, E.A.Puganova, et al. Analytical Chemistry 76 (2004) 474.
A.A.Karyakin, E.A.Puganova, I.A.Bolshakov, E.E.Karyakina, Angewandte Chemie-Int.
Ed. 46 (2007) 7678.
N.A.Sitnikova, A.V.Borisova, …, A.A.Karyakin Analytical Chemistry 83 (2011) 2359
•improved enzyme immobilization protocol
A. A. Karyakin, E. E. Karyakina, et. al. Analytical Chemistry 68 (1996) 4335.
A.A. Karyakin, E. A. Kotel'nikova et. al. Analytical Chemistry 74 (2002) 1597.
E.I. Yashina, A.V. Borisova, ... , A.A. Karyakin. Analytical Chemistry 82 (2010) 1601
A.N. Sekretaryova, …, A.A. Karyakin Analytical Chemistry 84 (2012) 1220
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Oxidase based biosensors
(> 90% of biosensors, kits, etc.)
Oxidase
Analyte
O2
Oxidized
Analyte
H2O2
Monitoring of H2O2 formation:
Advantages: -low detection limit (<1 M).
How to detect H2O2?
1. Pt electrode, +0.6 V, Ag/AgCl.(G.G. Guilbault, et al.. Anal.Chem. 45 (1973)2255)
2. Peroxidase electrode.
Disadvantages: reductants (ascorbate, paracetamol etc.) generate
parasitic signal
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Prussian Blue FeIII4[FeII(CN)6]3:
nano-scaled layer - advanced electrocatalyst
Selectivity
jH2O2/jO2
Electrochem.
constant,
cm/s
0.03
k, cm s
-1
Electrocatalyst
Pt
0.1
0.02
4.10-6
0.01
Prussian Blue
400-600
1.10-2
PB, nmol cm
-2
0.00
0
5
10
15
thickness 40-80 nm
A.A.Karyakin, E.E.Karyakina, L.Gorton. Analytical Chemistry 72 (2000) 1720
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Superstable H2O2 transducer
(continuous flow of 1 mM H2O2)
N.A. Sitnikova, A.V. Borisova, M.A. Komkova, A.A. Karyakin. Analytical Chemistry
83 (2011) 2359-63
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Nano-electrode arrays
signal:noise ~ 1/r
Nano-structuring of electrocatalyst
onto inert support
A.A. Karyakin, E.A. Puganova et.al. Analytical Chemistry, 2004, 76, 474-8
8
10
-2
10
-4
10
-6
15
i, nA cm
-2
i, A cm
-2
Electrochemical sensor with record
performance characteristics
10
10
-8
5
-1
0
10
-9
10
-8
10
-7
10
-6
10
[H2O2], nmol L
0
-5
5
10
-4
10
10
[H2O2], mol L
15
-3
10
20
-2
25
10
30
-1
-1
A.A. Karyakin, E.A. Puganova, I.A. Bolshakov, E.E. Karyakina
Angewandte Chemie 46 (2007) 7678-80.
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Improved enzyme
immobilization protocol
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Immobilization from waterorganic mixtures
Reproducibility
Uniform membranes
Mixing enzymes with waterinsoluble polymers
Deposition from real polymer
solution
Exposing enzymes to organic solvents
A.A.Karyakin, E.E.Karyakina, L.Gorton et.al. Anal. Chem. 68 (1996) 4335.
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Combination of transducer and immobilization
protocol – advanced biosensors
Sensitivity compared to
Biosen C_Line
Stability:
> 500 detections without calibration
5.10 M
-4
-2
-80
i, A cm
glucose
glutamate
lactate
-60
-40
1.10 M
-4
-20
5.10 M
-5
5.10 M
-6
100 times improved
1.10 M
-5
0
0
500
1000
1500
2000
время, с
A.A. Karyakin, E.E. Karyakina, et. al. Analytical Chemistry 68 (1996) 4335.
A.A. Karyakin, E.E.Karyakina, L.Gorton. Analytical Chemistry 72 (2000) 1720
A.A. Karyakin, E.A. Kotel'nikova et. al. Analytical Chemistry 74 (2002) 1597.
E.I. Yashina, A.V. Borisova, ... , A.A. Karyakin. Analytical Chemistry 82 (2010) 1601
12
Lactate in blood:
correlation with the standard method
[lactate], mM (Biosensor)
10.0
27 sportsmens before
and after exercise stress
7.5
r > 0.99
5.0
2.5
0.0
0.0
2.5
[lactate],
5.0
7.5
mM (Humastar 300)
10.0
Non-invasive diagnostics
Correlation in metabolite concentration
with blood is required
Suitable excretory liquids:
Exhaled breath condensate;
Sweat;
Tears.
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ta
s
ia
ia
B
ro
nc
h
ie
c
m
on
O
PD
C
Pn
eu
ro
H
ea
nc
lth
hi
a
y
lA
B
A,
st
ex
hm
ac
a
er
ba
B
tio
A,
n
re
m
is
si
on
5
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
B
[H2 O2 ],  М
Non-invasive diagnostics of oxidative
stress: condensate of exhaled breath
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Human stress control upon
physical
training
•Lactate detection in sweat
before
training
before
training
after
training
Member of national junior
skiing team
[lactate] (after/before), sweat
Universal sweat collector
after
training
4
2
0
0
2
4
[lactate] (after/before), blood
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