Early detection of cell activation by ATR-FTIR spectroscopy

Patent metadata

Field	Value
Publication number	US-9983129-B2
Application number	US-201415102091-A
Country	US
Kind code	B2
Filing date	Dec 4, 2014
Priority date	Dec 5, 2013
Publication date	May 29, 2018
Grant date	May 29, 2018

Abstract

Disclosed are methods, systems, and apparatuses for rapidly detecting a cellular interaction, such as ligand:receptor interactions. For example, the disclosed methods and systems can be used to detect a cellular interaction within 15 minutes to 75 minutes. This allows cells to be used as biosensors to detect cell activating agents in a sample.

First claim

What is claimed is:

1. A method for detecting a cellular response resulting from a defined treatment or exposure, comprising
(a) depositing a sample comprising a plurality of cells under reaction conditions on an internal reflection element (IRE);
(b) directing a beam of infrared (IR) radiation through the IRE under conditions such that the IR radiation interacts with the plurality of cells;
(c) recording an absorption spectrum over a range of preselected frequencies; and
(d) comparing the absorption spectrum to a control spectrum;
wherein a change in absorbance at one or more frequencies compared to the control spectrum is an indication of a cellular interaction in the plurality of cells; and
wherein the reaction conditions comprise contacting the plurality of cells with a sample comprising one or more potential pathogens, allergens, or ligands.

2. The method of claim 1 , wherein the potential pathogen is a virus, bacteria, or yeast.

3. The method of claim 1 , wherein the potential ligand is selected from the group consisting of antibodies, growth factors, cytokines, chemokines, hormones, extracellular matrix proteins, and cell-surface proteins.

4. The method of claim 1 , wherein the potential ligand is selected from the group consisting of proteins, peptides, peptide nucleic acids, and small molecules.

5. The method of claim 1 , wherein the reaction conditions further comprise a change in temperature, pH, salinity, or any combination thereof.

6. The method of claim 1 , wherein the cellular interaction is detected within 15 minutes to 75 minutes.

7. The method of claim 1 , wherein the plurality of cells comprise bacterial or yeast cells.

8. The method of claim 1 , wherein the plurality of cells comprise mammalian cells.

9. The method of claim 1 , wherein the plurality of cells comprise a transformed cell line.

10. The method of claim 1 , wherein the range of preselected frequencies is between 50 cm −1 and 3700 cm −1 .

11. The method of claim 10 , wherein the range of preselected frequencies is between 800 cm −1 and 1500 cm −1 .

12. The method of claim 1 , wherein the IRE is an attenuated total reflectance (ATR) crystal comprising an optical material with a higher refractive index than the sample comprising the plurality of cells.

13. The method of claim 12 , wherein the IRE comprises a germanium crystal or a zinc selenide crystal.

14. The method of claim 1 , wherein the IR radiation that interacts with the plurality of cells is an evanescent wave with an average penetration depth of about 2 μm.

15. The method of claim 1 , further comprising Fourier transformation of the absorbance spectrum.

16. A method for using cells as a biosensor, comprising:
(a) exposing a homogeneous population of cells with a sample;
(b) depositing the homogeneous population of cells on an internal reflection element (IRE);
(c) directing a beam of infrared (IR) radiation through the IRE under conditions such that the IR radiation interacts with the plurality of cells;
(d) recording an absorption spectrum over a range of preselected frequencies; and
(e) comparing the absorption spectrum to a control spectrum;
wherein a change in absorbance at one or more frequencies compared to the control spectrum is an indication of a cell activating agent in the sample; and
wherein the cell activating agent comprises one or more potential pathogens, allergens, or ligands.

17. A system for detecting a cell activating agent in a sample, comprising:
(a) a Fourier transform infrared spectrometer configured with an internal reflection element (IRE) for attenuated total reflectance (ATR); and
(b) a homogeneous population of cells selected to react with the cell activating agent; and
wherein the cell activating agent comprises one or more potential pathogens, allergens, or ligands.

Assignees

• Univ Georgia State Res Found

Inventors

• Titus Jitto
• Filfili Chadi
• Perera A G Unil
• Hilliard Julia K

Classifications

• B07C5/3416

  according to radiation transmissivity, e.g. for light, x-rays, particle radiation (detecting hidden objects G01V) · CPC title

• G01N21/35

  using infrared light (G01N21/39 takes precedence) · CPC title

• G01N21/552 Primary

  Attenuated total reflection · CPC title

• G01N2021/3595

  using FTIR · CPC title

• G01N33/483

  Physical analysis of biological material · CPC title