Metal-antibody tagging and plasma-based detection

Patent metadata

Field	Value
Publication number	US-10514338-B2
Application number	US-201615042969-A
Country	US
Kind code	B2
Filing date	Feb 12, 2016
Priority date	Sep 12, 2014
Publication date	Dec 24, 2019
Grant date	Dec 24, 2019

Abstract

A target within a sample can be characterized using an energy source configured to transform a metal in the sample into a plasma and an optical spectroscopic detector configured to detect electromagnetic radiation emitted by the plasma to provide an optical-spectrum signal. A processor can determine presence of the metal in the sample using the optical-spectrum signal. The target can include a microbe or biological toxin. A recognition construct comprising a metal and a scaffold can be applied to the sample. The scaffold can bind to the target. Energy can be applied to transform at least some of the sample into a plasma. Electromagnetic radiation emitted by the plasma can be detected to provide an optical-spectrum signal of the sample. A preparation subsystem can add the recognition construct to the sample and a washing subsystem can wash unbound recognition construct out of the sample.

First claim

The invention claimed is:

1. A system for characterizing a target within a sample, the system comprising:
a substrate configured to retain the sample, the substrate comprising a silicon surface and a capture antibody coupled to the silicon surface, the capture antibody being configured to attach to the target;
the sample comprising a biomolecular recognition construct, wherein the biomolecular recognition construct comprises:
a scaffold configured to couple with the target;
a polymer coupled to the scaffold and comprising a metal-chelating ligand; and
a first metal atom or ion linked to the metal-chelating ligand;

an energy source configured to transform the first metal atom or ion in the sample into a plasma;
an optical spectroscopic detector configured to detect electromagnetic radiation emitted by the plasma and to provide an optical-spectrum signal of the sample based at least in part on at least some of the electromagnetic radiation;
a processor; and
a memory storing instructions executable by the processor to cause the processor to perform operations comprising:
generating a normalized optical-spectrum signal by normalizing the optical-spectrum signal of the sample with respect to an optical-spectrum signal of a material of the substrate; and
determining presence of the first metal atom or ion in the sample based at least in part on the normalized optical-spectrum signal.

2. The system according to claim 1 , the operations further comprising:
performing spectral unmixing on the optical-spectrum signal of the sample;
performing constrained energy minimization on the optical-spectrum signal of the sample; or
determining presence of the metal atom or ion in the sample by applying the optical-spectrum signal of the sample to a multi-class classifier selected from the group consisting of a support vector machine, a kernel estimator, a nearest-neighbor classifier, a decision tree, a decision forest, a neural network, or a deep neural network.

3. The system according to claim 1 , the operations further comprising:
determining presence of a second metal atom or ion in the sample based at least in part on the optical-spectrum signal of the sample, wherein:
the first metal atom or ion is an atom or ion of a first element; and
the second metal atom or ion is an atom or ion of a second element different from the first element.

4. The system according to claim 1 , wherein the substrate comprises a silicon wafer.

5. The system according to claim 1 , wherein the substrate comprises recognition macromolecules that comprise the capture antibody.

6. The system according to claim 1 , wherein the scaffold comprises at least one of: adNectin, iMab, anticalin, designed ankyrin repeat protein (DARPin), affilin, tetranectin, or avimer.

7. The system according to claim 1 , wherein the biomolecular recognition construct consists essentially of:
the scaffold;
the polymer; and
the first metal atom or ion bound to the metal-chelating ligand.

8. The system according to claim 7 , wherein the scaffold consists essentially of a structure selected from the group consisting of adNectin, iMab, anticalin, designed ankyrin repeat protein (DARPin), affilin, tetranectin, and avimer.

9. The system according to claim 1 , wherein the biomolecular recognition construct comprises:
at least one scaffold, the at least one scaffold comprising the scaffold;
one or more polymers, the one or more polymers comprising the polymer; and
one or more metal atoms or ions, each of the one or more metal atoms or ions bound to a respective metal-chelating ligand of a corresponding polymer of the one or more polymers, wherein the one or more metal atoms or ions comprise the first metal atom or ion, and the metal-chelating ligand is one of the respective metal-chelating ligands.

10. The system according to claim 9 , wherein the scaffold consists of a structure selected from the group consisting of adNectin, iMab, anticalin, designed ankyrin repeat protein (DARPin), affilin, tetranectin, and avimer.

11. The system according to claim 1 , wherein the metal-chelating ligand comprises diethyl enetriaminepenta-acetic acid (DTPA),
wherein the first metal atom or ion linked to the metal-chelating ligand comprises a lanthanide.

12. The system according to claim 1 , wherein the capture antibody is an antitoxin antibody.

13. The system according to claim 1 , wherein generating the normalized optical-spectrum signal comprises normalizing the optical-spectrum signal of the sample with respect to an optical-spectrum signal of silicon.

Assignees

• Purdue Research Foundation

Inventors

• Robinson Joseph Paul
• Rajwa Bartlomiej P
• Patsekin Valery P
• Bae Euiwon

Classifications

• G01N21/67

  using electric arcs or discharges · CPC title

• G01N21/718 Primary

  Laser microanalysis, i.e. with formation of sample plasma · CPC title

• G01N33/58

  involving labelled substances (G01N33/53 takes precedence) · CPC title

• G01N33/56911

  Bacteria · CPC title

• G01N2469/00

  Immunoassays for the detection of microorganisms · CPC title