Real time electronic cell sensing systems and applications for cell-based assays

Patent metadata

Field	Value
Publication number	US-11604197-B2
Application number	US-202016833180-A
Country	US
Kind code	B2
Filing date	Mar 27, 2020
Priority date	Dec 20, 2002
Publication date	Mar 14, 2023
Grant date	Mar 14, 2023

Abstract

Methods for identifying compounds that modulate cellular responses stimulated by IgE, which include providing an impedance-based system that monitors cell-substrate impedance of cells on a substrate; introducing cells to the substrate of the system; adding at least one test compound and IgE to the cells, wherein the at least one test compound is suspected of modulating cell responses stimulated by the IgE; adding an antigen to the cells; monitoring the cell-substrate impedance of cells on the substrate; and analyzing the cell-substrate impedance to evaluate whether the at least one test compound alters a cellular response to stimulation with the IgE.

First claim

What is claimed is:

1. A method for identifying a target for modulating cellular responses to IgE stimulation, the method comprising:
a) providing an impedance-based system that monitors cell-substrate impedance of cells on a substrate;
b) introducing cells into the system, wherein the cells are genetically modified to alter a function of, or reduce or ablate expression of, a suspected target molecule;
c) adding IgE to the cells;
d) adding an antigen to the cells;
e) monitoring cell-substrate impedance of the cells on the substrate;
f) analyzing cell-substrate impedance data compared to control cells that are not genetically modified to identify a target for modulation of cellular responses to IgE stimulation.

2. The method of claim 1 , wherein the cells are primary cells.

3. The method of claim 1 , wherein the cells are selected from the group consisting of mast cells, eosinophils, basophils, and genetically engineered cells.

4. The method of claim 1 , wherein one or more compounds for genetically modifying the cells are added to the cells prior to the step of adding IgE.

5. The method of claim 1 , wherein cell-substrate impedance is monitored before the step of adding the IgE to the cells.

6. The method of claim 1 , wherein the cell-substrate impedance is monitored after the step of adding the IgE to the cells.

7. The method of claim 1 , wherein the cell-substrate impedance is monitored before and after the step of adding the IgE to the cells.

8. The method of claim 7 , wherein said cell-substrate impedance is monitored before and after the step of adding the antigen to the cells.

9. A method for comparing responses to IgE stimulation between cells of different genotypes, the method comprising:
a) providing an impedance-based system that monitors cell-substrate impedance of cells on a substrate;
b) introducing cells of two different genotypes to the substrate of the system to form two different genotype cell populations;
c) adding IgE to the cells of both genotypes;
d) adding an antigen to the cells of both genotypes;
e) monitoring cell-substrate impedance of the cells of both genotypes on the substrate; and
f) comparing the cell-substrate impedances between the cells of different genotypes.

10. The method of claim 9 , wherein the cells of two different genotypes are primary cells from human subjects.

11. The method of claim 9 , wherein the antigen is a known or suspected allergen.

12. The method of claim 9 , further comprising correlating the cell-substrate impedances with molecular genetic data obtained from the different genotypes.

13. The method of claim 12 , wherein the molecular genetic data is selected from one or more of the group consisting of RFLIP data, SNP data, DNA marker data, RNA expression profiling data, protein expression profiling data, and alternative splicing data.

14. A method for identifying an antigen or half antigen that can cause an allergic response, the method comprising:
a) providing an impedance-based system that monitors cell-substrate impedance of cells on a substrate;
b) introducing cells to the substrate of the system;
c) adding IgE to the cells;
d) adding at least one compound suspected of being an antigen or half antigen to the cells;
e) monitoring cell-substrate impedance of cells on the substrate to obtain cell-substrate impedance data of the cells treated with the antigen or the half antigen; and
f) analyzing the data to identify whether the compound is an antigen or half antigen.

15. The method of claim 14 , wherein the substrate is positioned in an incubator.

Assignees

• Agilent Technologies Inc

Inventors

• Xu Xiao
• Abassi Yama A
• Wang Xiaobo
• Gan Jiangbo

Classifications

• G01N33/6854 Primary

  Immunoglobulins · CPC title

• C12M41/36

  of biomass, e.g. colony counters or by turbidity measurements (electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404) · CPC title

• G01N33/4836

  using multielectrode arrays · CPC title

• G01N33/5005

  involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025) · CPC title

• G01N2500/10

  involving cells · CPC title