ECL-BASED ELECTRODE, MANUFACTURING METHOD FOR ECL-BASED ELECTRODE, ECL SENSOR, MANUFACTURING METHOD FOR ECL SENSOR, METHOD FOR ECL DETECTION OF NUCLEIC ACID-SPECIFIC SITE MODIFICATION, AND KIT FOR NUCLEIC ACID MODIFICATION DETECTION USED FOR METHOD FOR ECL DETECTION OF NUCLEIC ACID-SPECIFIC SITE MODIFICATION
A manufacturing method for an ECL (Electrochemiluminescence)-based electrode according to an embodiment includes a step of mixing an electrocatalytic solution and metal nanoparticles and thereafter dropping a mixed particle suspension onto the surface of an electrode and obtaining a base electrode co-modified by an electrocatalyst-metal nanoparticle.
1 . A manufacturing method for an ECL (electrochemiluminescence)-based electrode, comprising a step of mixing an electrocatalytic solution and metal nanoparticles and thereafter dropping a mixed particle suspension onto a surface of an electrode and obtaining a base electrode co-modified by an electrocatalyst-metal nanoparticles.
2 . The manufacturing method according to claim 1 ,
wherein the electrocatalyst is a transition metal compound.
3 . The manufacturing method according to claim 1 ,
wherein the electrocatalyst is molybdenum diselenide (MoSe 2 ).
4 . The manufacturing method according to claim 1 ,
wherein the metal nanoparticles are at least one of gold (Au) nanoparticles, silver (Ag) nanoparticles, platinum (Pt) nanoparticles, copper (Cu) nanoparticles, cobalt (Co) nanoparticles, iron (Fe) nanoparticles, nickel (Ni) nanoparticles, and multi-alloy nanoparticles thereof.
5 . The manufacturing method according to claim 1 ,
wherein the metal nanoparticles are gold (Au) nanoparticles.
6 . The manufacturing method according to claim 1 ,
wherein the electrode is one of a glass carbon electrode, an ITO (Indium Tin Oxide) electrode, and a screen printing electrode.
7 . The manufacturing method according to claim 1 ,
wherein the electrode is a glass carbon electrode.
8 . The manufacturing method according to claim 1 ,
wherein an exfoliation time of the electrocatalyst is five hours or longer.
9 . The manufacturing method according to claim 1 ,
wherein an exfoliation time of the electrocatalyst is five hours or longer and shorter than 25 hours.
10 . The manufacturing method according to claim 1 ,
wherein a mixing ratio of the electrocatalyst and the metal nanoparticles is 1:1 to 1:9 in a volume ratio of 3.5 mg/mL of MoSe 2 and 0.5 mg/mL of AuNPs.
11 . The manufacturing method according to claim 1 ,
wherein a mixing ratio of the electrocatalyst and the metal nanoparticles is 1:5 to 1:7 in a volume ratio of 3.5 mg/mL of MoSe 2 and 0.5 mg/mL of AuNPs.
12 . An ECL-based electrode comprising
an interface co-modified by an electrocatalytic solution and metal nanoparticles.
13 . A manufacturing method for an ECL sensor, comprising:
manufacturing an ECL-based electrode, with a manufacturing method for an ECL-based electrode including a step of mixing an electrocatalytic solution and metal nanoparticles and thereafter dropping a mixed particle suspension onto a surface of an electrode and obtaining a base electrode co-modified by an electrocatalyst-metal nanoparticles; and
a step of binding a terminal-modified capture nucleic acid to a surface of the base electrode.
14 . The manufacturing method according to claim 13 ,
wherein the terminal-modified capture nucleic acid is a mercapto-modified capture nucleic acid, an amino-modified capture nucleic acid, or a biotin-modified capture nucleic acid.
15 . The manufacturing method according to claim 14 ,
wherein the terminal-modified capture nucleic acid is a mercapto-modified capture nucleic acid.
16 . The manufacturing method according to claim 13 ,
further comprising a step of blocking, with a blocking agent, the surface of the electrode to which the capture nucleic acid is bound.
17 . The manufacturing method according to claim 16 ,
wherein the blocking agent is at least one of a small molecule blocking agent and a protein blocking agent.
18 . The manufacturing method according to claim 17 ,
wherein the small molecule blocking agent is 6-mercapto-1-hexanol (MCH).
19 . The manufacturing method according to claim 17 ,
wherein the protein blocking agent is bovine serum albumin.
20 . An ECL sensor comprising:
an ECL-based electrode having an interface co-modified by an electrocatalytic solution and metal nanoparticles; and
a terminal-modified capture nucleic acid bound to a surface of the ECL-based electrode.
21 . An ECL detection method for nucleic acid-specific site modification, comprising:
a first step of dropping a test sample onto a surface of an ECL sensor including an ECL-based electrode having an interface co-modified by an electrocatalytic solution and metal nanoparticles and a terminal-modified capture nucleic acid bound to a surface of the ECL-based electrode and capturing, with the capture nucleic acid, a target nucleic acid on the surface of the ECL sensor;
a second step of dropping an anti-nucleic acid modification antibody and binding the anti-nucleic acid modification antibody to a nucleic acid-specific site on the target nucleic acid;
a third step of adding an ECL nanoprobe and labeling the target nucleic acid with a detection signal, and
a fourth step of immersing the ECL sensor obtained in the third step in a detection solution containing a co-reactive agent to measure an ECL signal.
22 . The detection method according to claim 21 , wherein
the ECL nanoprobe is metal-doped inorganic oxide nanoparticles modified by a secondary antibody, and
the secondary antibody is affinity-bound to the anti-nucleic acid modification antibody.
23 . The detection method according to claim 22 ,
wherein the inorganic oxide nanoparticles are nanoparticles coated with silicon dioxide, titanium dioxide, zinc oxide, or iron oxide, silicon dioxide nanoparticles, titanium dioxide nanoparticles, zinc oxide nanoparticles, or iron oxide nanoparticles.
24 . The detection method according to claim 22 ,
wherein the inorganic oxide nanoparticles are silicon dioxide nanoparticles.
25 . The detection method according to claim 22 ,
wherein the metal-doped inorganic oxide nanoparticles are silicon dioxide nanoparticles in which the metal-doped inorganic oxide nanoparticles are doped with tris(bipyridine) ruthenium(II) complex ions (Ru(bpy) 3 2+ ).
26 . The detection method according to claim 21 ,
wherein the co-reactive agent is tripropylamine.
27 . The detection method according to claim 21 ,
wherein the nucleic acid modification is methylation modification, methylolation modification, or formylation modification of a nucleic acid.
28 . The detection method according to claim 21 , wherein
pH of the detection solution is 6.0 or higher,
concentration of the capture nucleic acid is 1 nM to 400 nM,
an incubation time of the capture nucleic acid is five minutes or longer,
an incubation time of the target nucleic acid is five minutes or longer,
concentration of the anti-nucleic acid modification antibody is 1 μg/mL or higher,
an incubation time of the anti-nucleic acid modification antibody is five minutes or longer,
concentration of the ECL nanoprobe is 1 μg/mL or higher, and
an incubation time of the ECL nanoprobe is five minutes or longer.
29 . The detection method according to claim 21 , wherein
pH of the detection solution is 6.5 to 8.5,
concentration of the capture nucleic acid is 20 nM to 100 nM,
an incubation time of the capture nucleic acid is 10 minutes or longer,
an incubation time of the target nucleic acid is 10 minutes or longer,
concentration of the anti-nucleic acid modification antibody is 2 μg/mL or higher,
an incubation time of the anti-nucleic acid modification antibody is 10 minutes or longer,
concentration of the ECL nanoprobe is 2 μg/mL or higher, and
an incubation time of the ECL nanoprobe is 10 minutes or longer.
30 . A kit for nucleic acid modification detection used in an ECL detection method for nucleic acid-specific site modification, comprising:
an ECL-based electrode having an interface co-modified by an electrocatalytic solution and metal nanoparticles;
a terminal-modified capture nucleic acid bound to a surface of the ECL-based electrode;
an anti-nucleic acid modification antibody;
an ECL nanoprobe; and
a detection solution.
31 . An ECL-based electrode,
co-modified by an electrocatalyst and metal nanoparticles.
32 . The ECL-based electrode according to claim 31 ,
wherein the ECL-based electrode does not contain graphene.