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!"#$%&'#()*+,--%.%*/!0-#"***!***/!0-#"*+#()*!"#$%&'#()***!***()"/1--*"1(#,"1-*#"'(#(!(%**
Vincenzo F. Curto
Ionic Liquids in biosensors: influence of
hydrated choline based ILs on the
bio-functionality of Lactate Oxidase
!"#$%&'#()*+,--%.%*/!0-#"***!***/!0-#"*+#()*!"#$%&'#()***!***()"/1--*"1(#,"1-*#"'(#(!(%** 2
Electrochemical and wearable biosensors
Long-life bioreceptor issues
" narrow pH range activity
" temperature range stability
" limitations that are associated with the use of conventional aqueous electrolytes
!
Wearable sensors allow the continuous monitoring of a person’s physiology in a natural setting
!"#$%&'#()*+,--%.%*/!0-#"***!***/!0-#"*+#()*!"#$%&'#()***!***()"/1--*"1(#,"1-*#"'(#(!(%**
Ionogel & OECTs: Lactate Sensor
3
V. F. Curto, et al., J. Mater. Chem., 22, 4440 (2012).
!"#$%&'#()*+,--%.%*/!0-#"***!***/!0-#"*+#()*!"#$%&'#()***!***()"/1--*"1(#,"1-*#"'(#(!(%**
Enzyme: Lactate Oxidase (LOx) from Aerococcus Viridans
– !-hydroxy-acid oxidase flavoenzyme family
– Molecular Weight: 80 kDa
Choline based ILs
1. Choline aminoacetate
2. Choline dhp
3. Choline levulinate 4. Choline bis(2-ethylhexyl)phosphate (BEH)
5. Choline formate
6. Choline tartrate
7. Choline gallate
8. Choline dibutylphosphate
9. Choline valporate
10. Choline nitrate
11. Choline chloride
4
!"#$%&'#()*+,--%.%*/!0-#"***!***/!0-#"*+#()*!"#$%&'#()***!***()"/1--*"1(#,"1-*#"'(#(!(%**
234444*
25444*
26444*
27444*
28444*
4*
8444*
7444*
6444*
5444*
34444*
38444*
37444*
36444*
35444*
394* 844* 834* 884* 8:4* 874* 8;4*
!"#$%&"'()*
"%"++(,
-.(/0
%1)"2%.!
3 %)!4+
567%
%
8#9"+"$2/:%1$!7%
8;<+*=>?@A>*B>CDEF*
9;<+*
8;<+*CG>A*B>CDEF*
2;444*
27444*
2:444*
28444*
23444*
4*
3444*
8444*
:444*
7444*
;444*
6444*
H444*
394* 844* 834* 884* 8:4* 874* 8;4*
!"#$%&"'()*
"%"++(,
-.(/0
%1)"2%.!
3 %)!4+
567%
%
8#9"+"$2/:%1$!7%
8;*<+*=>?@A>*B>CDEF*
9;<+*
8;<+*CG>A*B>CDEF*
Enzyme structure - Cirlcular Dichroism
PBS Choline dhp
5
!"#$%&'#()*+,--%.%*/!0-#"***!***/!0-#"*+#()*!"#$%&'#()***!***()"/1--*"1(#,"1-*#"'(#(!(%**
0
20
40
60
0 1.5 3 4.5 6
Velo
city
[U m
g-1 ]
Concentration [mM]
IL free Choline Cl choline dhp choline formate Choline nitrate Choline levulinate
Enzyme Kinetic
6
Km [mM] ;<%=&""% 0.366
>:4+($"%>+% 0.377 >:4+($"%):,% 0.825
Km [mM] >:4+($"%=4&!#/"% 0.926 >:4+($"%$(/&#/"% 0.487
>:4+($"%+"9*+($#/"% 0.674
!"#$%&'#()*+,--%.%*/!0-#"***!***/!0-#"*+#()*!"#$%&'#()***!***()"/1--*"1(#,"1-*#"'(#(!(%**
0
25
50
75
100
0 30 60 90 120 150
Nor
mal
ised
enz
yme
velo
city
(v/v
o) %
time/day
#-*?A>>*
+B@IJE>*+I*
+B@IJE>*KBL*
+B@IJE>*"JMACM>*
+B@IJE>*?@ANCM>*
+B@IJE>*MCAMACM>*
+B@IJE>*I>OPIJECM>*
+B@IJE>*KJ=PMQILB@RLBCM>*
Long Term stability – T= 5 °C
7
!"#$%&'#()*+,--%.%*/!0-#"***!***/!0-#"*+#()*!"#$%&'#()***!***()"/1--*"1(#,"1-*#"'(#(!(%**
Dr. Dion Khodagholy Prof. Róisín Owens Prof. George Malliaras
Dr. Fernando Benito-Lopez Prof. Dermot Diamond
Dr. Vijay Ranganathan Prof. Doug MacFarlane
Acknowledgements
07/CE/I1147
Research Career Start Programme 2010