Granted Patent B2

US 12,292,407 · App. 17/184,274 · Granted May 6, 2025

Inventors: Young Shik Shin (Mountain View, CA); Nadezda Fomina (Redwood City, CA); Christopher Johnson (San Carlos, CA); Armin Darvish (Brisbane, CA); Christoph Lang (Sunnyvale, CA)

Assignee: Robert Bosch GmbH

G01N27/4167C12Q1/6851G05D21/02

Abstract

Claims (45)

1. A method for monitoring a chemical reaction, comprising

(a) starting the chemical reaction in a buffered solution equipped with a closed-loop device, wherein

the buffered solution comprises a pH modulating agent;

the closed-loop device comprises an electronic controller, a working electrode, a counter electrode, a reference electrode, and a sensing element, wherein the working electrode, the counter electrode, the reference electrode, and the sensing element are immersed in the buffered solution, and wherein the sensing element is positioned on top of the working electrode with an insulation layer therebetween to provide physical separation; and

the chemical reaction causes a change of ionic strength of the buffered solution;

(b) applying a current or voltage to the working electrode, whereupon the pH modulating agent causes a continuous cycling of pH modulation between at least two target pH values in a zone adjacent to a surface of the working electrode and the sensing element;

(c) detecting the change of ionic strength of the buffered solution with the closed-loop device, wherein the detecting comprises:

(c 1 ) controlling the current or voltage applied to the working electrode, thereby modulating the pH value in the zone between the at least two target pH values;

(c 2 ) analyzing the current or voltage applied to the working electrode and a signal output from the sensing element; and

(c 3 ) quantitating the change of ionic strength of the buffered solution from the current or voltage applied to the working electrode and the signal output from the sensing element; and

(d) repeating steps (b) and (c) at least one time.

2. The method of claim 1 , wherein detecting further comprises analyzing one or more additional parameters of the closed-loop device selected from the group consisting of: a first target pH value, a second target pH value, pulse duration at the first target pH value (ts 1 ), pause duration at the second target pH value (ts 2 ), pH rise time (t 1 ), pH decay time (t 2 ); rise peak current (c 1 ), stable current (c 2 ), decay peak current (c 3 ), current rise time (t 3 ), and current decay time (t 4 ).

3. The method of claim 1 , further comprising

(e) determining the progress of the chemical reaction.

4. The method of claim 1 , wherein the sensing element comprises

an ion-sensitive field-effect transistor (ISFET);

a metal oxide electrode selected from the group consisting of thallium oxide electrode, aluminum oxide electrode, tin oxide electrode, zinc oxide electrode, ruthenium oxide electrode, titanium dioxide electrode, and iridium oxide electrode; or

an electrode coated with a pH sensitive coating, the pH sensitive coating comprising a material selected from the group consisting of polyaniline, polypyrrole, polyaminoanthracene, polycarbazole, polybisphenol A, polyethyleneimine, poly (p-phenylenediamine), poly-1,5- diaminonaphthalene, titanium nitride, thallium oxide, aluminum oxide, tin oxide, zinc oxide, ruthenium oxide, and iridium oxide.

5. The method of claim 1 , wherein a bulk of the buffered solution apart from the zone adjacent to the surface of the working electrode maintains a constant pH value.

6. The method of claim 1 , wherein the buffered solution comprises a buffering agent selected from the group consisting of phosphate, TAPS, Bicine, Tris, Tricine, TAPSO, HEPES, TES, MOPS, PIPES, cacodylate, SSC, and MES.

7. The method of claim 6 , wherein a concentration of the buffering agent in the buffered solution is at least 1 mM.

8. The method of claim 1 , wherein the chemical reaction produces H + or OH 31 .

9. The method of claim 1 , wherein the chemical reaction is a DNA amplification reaction.

10. The method of claim 1 , wherein the chemical reaction is a quantitative Polymerase Chain Reaction (qPCR).

11. The method of claim 1 , wherein the pH modulating agent is a quinone derivative, an aminophenol derivative, an aniline derivative, a benzidine derivative, a hydrazine derivative, or a combination thereof.

12. The method of claim 1 , wherein the pH modulating agent is a quinone derivative of any of formula (I)-(XII)

wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8 are each independently selected from the group consisting of: H; C n H 2n+1 ; Cl; F; I, Br, OM, NO 2 , OH, OC n H 2n+1 , OC n H 2n OH, O(C n H 2n O) y H,

O(C n H 2n O) y C n H 2n+1 , O(C n H 2n O) y COOH; O(C n H 2n O) y COOM; COOH; COOM; COOC n H 2n+1 ;

CONHC n H 2n+1 ; CON(C n H 2n+1 ) 2 ; SO 3 H; SO 3 M; NH 2 ; NHC n H 2n+1 ; N(C n H 2n+1 ) 2 ; NHC n H 2n OH;

NHC n H 2n NH 2 ; N(C n H 2n OH) 2 ; N(C n H 2n NH 2 ) 2 ; NHCOC n H 2n+1 ; NC n H 2n COC n H 2n+1 ;

NC n H 2n COC n H 2n OH; NC n H 2n COC n H 2n NH 2 ; NHC n H 2n COC n H 2n SH; SH; SC n H 2n+1 ; SC n H 2n OH;

S(C n H 2n O) y H; S(C n H 2n O) y C n H 2n+1 ; S(C n H 2n O) y COOH; S(C n H 2n O) y COOM; OC n H 2n SH;

O(C n H 2n O) y C n H 2n SH; O(C n H 2n O) y C n H 2n SC n H 2n+1 ; C n H 2n OC n H 2n+1 ; C n H 2n SC n H 2n+1 ;

C n H 2n NHC n H 2n+1 ; C n H 2n OH; C n H 2n OC n H 2n+1 ; C n H 2n OC n H 2n OH; C n H 2n O(C n H 2n O) y COOH;

C n H 2n O(C n H 2n O) y COOM; C n H 2n COOH; C n H 2n COOM; C n H 2n COOC n H 2n+1 ;

C n H 2n CONHC n H 2n+1 ; C n H 2n CONH(C n H 2n+1 ) 2 ; C n H 2n SO 3 H; C n H 2n SO 3 M; C n H 2n NH 2 ;

C n H 2n N(C n H 2n+1 ) 2 ; C n H 2n NHC n H 2n OH; C n H 2n NHC n H 2n NH 2 ; C n H 2n N(C n H 2n OH) 2 ;

C n H 2n N(C n H 2n NH 2 ) 2 ; C n H 2n NHCOC n H 2n+1 ; C n H 2n NHC n H 2n COC n H 2n OH;

C n H 2n NHC n H 2n COC n H 2n NH 2 ; C n H 2n NHC n H 2n COC n H 2 SH; C n H 2n SH; C n H 2n SC n H 2n+1 ;

C n H 2 SC n H 2n OH; C n H 2n S(C n H 2n O) y H; C n H 2n S(C n H 2n O) y C n H 2n+1 ; C n H 2n S(C n H 2n O) y C n H 2n COOH;

C n H 2n S(C n H 2n O) y C n H 2n COOM; sugars; peptides; and amino acids,

wherein

M is any metal cation or NH 4 + ,

n is an integer from 1 to 10 9 , and

y is an integer from 1 to 10 9 .

Assignments (1)

Continuity (1)

References Cited (26)