Mass tag analysis for rare cells and cell free molecules

Patent metadata

Field	Value
Publication number	US-12480942-B2
Application number	US-202218075578-A
Country	US
Kind code	B2
Filing date	Dec 6, 2022
Priority date	Sep 24, 2015
Publication date	Nov 25, 2025
Grant date	Nov 25, 2025

Abstract

The invention generally relates to mass tag analysis for rare cells and cell free molecules. In certain embodiments, the invention provides an apparatus including an essentially non-absorbent membrane having at least one pore, a microwell operably associated with the essentially non-absorbent membrane, and an electric field generator. The apparatus may be configured such that an electric field produced by the electric field generator operably interacts with a sample in the microwell and expels a droplet of the sample through the at least one pore in the essentially non-absorbent membrane. In certain embodiments, apparatuses of the invention are used for detection, and optionally quantification, of a target analyte from a heterogeneous sample, such as a rare target analyte (e.g., rare cell) from a biological sample.

First claim

What is claimed is:

1 . A method for detecting a target analyte from a heterogeneous sample, the method comprising:
providing an apparatus that includes an essentially non-absorbent membrane comprising at least one pore, a microwell operably associated with the membrane, and an electric field generator operably associated with the membrane; wherein the microwell comprises a back wall, side walls, and a front that is operably associated with the essentially non-absorbent membrane, wherein the back wall, the side walls, and the front define a volume within the microwell; and wherein the electric field generator is physically separate and spaced apart from the back wall and side walls of the microwell and positioned within a space behind the back wall of the microwell and aligned within the space to not be outside of the side walls of the microwell such that the electric field generator inductively imparts an electric field to a sample within the volume within the microwell defined by the back wall, the side walls, and, and the front of the microwell to operably interact with the sample in the volume within the microwell and expel a droplet of the sample through the at least one pore in the essentially non-absorbent membrane; introducing a heterogeneous sample to at least the membrane; introducing to the sample a plurality of affinity agents that each comprise a first molecule, wherein the plurality of affinity agents specifically bind the target analyte in the sample; removing unbound affinity agents; introducing one or more additional molecules to the sample, wherein the one or more additional molecules interact with the first molecule to form a mass spectrometry label; providing voltage to the sample via the electric field generator to release a droplet through the at least one pore, wherein the droplet comprises a portion of the sample and the mass spectrometry label; and analyzing the droplet for presence of the mass spectrometry label, wherein the presence of the mass spectrometry label indicates presence of the target analyte in the sample.

2 . The method according to claim 1 , wherein the first molecule is a mass spectrometry label precursor.

3 . The method according to claim 2 , wherein the one or more additional molecules is an alteration agent that interacts with the mass spectrometry label precursor to form the mass spectrometry label.

4 . The method according to claim 1 , wherein the affinity agent is a particulate.

5 . The method according to claim 1 , wherein the target analyte is a rare cell and the heterogeneous sample is a heterogeneous biological sample.

6 . The method according to claim 1 , further comprising quantifying the target analyte in the sample by quantifying an amount of mass spectrometry label analyzed.

Assignees

• Purdue Research Foundation

Inventors

• Baird Zane
• Cooks Robert Graham
• Hollerbach Adam
• Ouyang Zheng
• Pugia Michael

Classifications

• G01N2458/15

  Non-radioactive isotope labels, e.g. for detection by mass spectrometry · CPC title

• G01N33/5302 Primary

  Apparatus specially adapted for immunological test procedures · 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

• G01N33/6848 Primary

  Methods of protein analysis involving mass spectrometry · CPC title