Biomedical Engineering Reference
In-Depth Information
3
2
1
a
b
1000
0 min
100
10
1000
10 min
100
10
1000
20 min
100
10
60 min
1000
100
10
18
12
60
-
-
-
2 s
I (pA)
Fig. 13.9 Monitoring of A-
(10-20) cleavage by trypsin. (a) Representative segments of a single
channel recording trace at various times.
Dashed lines
represent the levels of zero current; (b) The
corresponding time-dependent event amplitude histograms. Dashed lines 1, 2, and 3 represent the
mean residual current levels for peptides YEVHHQKLVFF, YEVHHQK, and LVFF, respectively.
The experiment was performed at
b
M trypsin. Figure
reprinted with permission from [
61
], Copyright 2009 American Chemical Society
40 mV with 10
m
MA-
b
(10-20) and 0.025
m
(10-20) were clearly observed. The identities of these two new types of events,
which were believed to be attributed to the two breakdown products of A-
b
(10-20), were further confirmed by investigating the translocation of individual
peptide standards. It is worth mentioning that in addition to the determination of the
identities of short peptide fragments, this approach could be also employed as a
real-time label-free method to study enzyme kinetics, e.g., to obtain
V
max
and
K
m
.
This may be very useful since incorporation of a label may alter the structure and
function of the target biomolecule.
13.4.2 DNA
Nanopore DNA sequencing, first introduced in the mid-1990 s [
17
], has emerged as
one of the most promising sequencing technology among the other sequencing
methods [
62
,
63
]. A good overview of the current status of nanopore DNA
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