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Personal glucose meters for detection and quantification of a broad range of analytes

A technology of glucose and substances, applied in the field of sensors, can solve the problem of not being able to provide quantitative results, etc.

Active Publication Date: 2013-04-03
THE BOARD OF TRUSTEES OF THE UNIV OF ILLINOIS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these sensors are qualitative or semi-quantitative and thus cannot provide quantitative results

Method used

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  • Personal glucose meters for detection and quantification of a broad range of analytes
  • Personal glucose meters for detection and quantification of a broad range of analytes
  • Personal glucose meters for detection and quantification of a broad range of analytes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0232] Materials and methods

[0233] Streptavidin-coated magnetic beads, PD-10 size exclusion columns, and Amicon-100K centrifugal filters were purchased from Bangs Laboratories Inc. (Fishers, IN), GE Healthcare Life Science Ltd. (Piscataway, NJ) and Millipore, respectively. Inc. (Billerica, MA). Invertase from class VII baker's yeast (S. cerevisiae), human recombinant interferon-gamma (IFN-gamma), 4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid sulfonic acid succinimidyl ester (sulfo-SMCC), 1,4-phenylene diisothiocyanate (PDITC), tris(2-carboxyethyl)phosphine hydrochloride (TCEP) and others used in Buffer chemicals and solvents were purchased from Sigma-Aldrich Inc. (St. Louis, MO). The following oligonucleotides were purchased from Integrated DNA Technologies Inc. (Coralville, IA):

[0234] Biotin-modified DNA (biotin-DNA):

[0235] TCACAGATGAGTAAAAAAAAAAAAA-biotin' (SEQ ID NO: 1)

[0236] Thiol-modified DNA (thiol-DNA):

[0237] HS-AAAAAAAAAAAAAGTCTCCCGAGAT-FAM' (...

Embodiment 2

[0279] Conjugation of invertase to ssDNA

[0280] This example describes a method for conjugating invertase to single-stranded (ss) DNA. Those skilled in the art will understand that other enzymes that catalyze the conversion of substances to glucose, such as cellulase, which converts cellulose to glucose, may be used in place of the invertase.

[0281] Invertase (from baker's yeast), also known as beta-fructofuranosidase, is an enzyme that catalyzes the hydrolysis of sucrose 50 . It can be used for signal amplification because nanomolar levels of the enzyme are capable of efficiently converting up to millimolar levels of sucrose to fructose and glucose at room temperature within a reasonable length of time and does not require laboratory-based setups. Additionally, only the fructose and glucose produced can be detected using the widely available personal glucose meter (PGM), whereas sucrose is completely "silent" in the PGM, not producing any signal or interference due to i...

Embodiment 3

[0285] Immobilization of functional DNA and DNA-invertase conjugates to magnetic beads

[0286] This example describes the method for immobilizing the invertase-ssDNA molecules generated in Example 2 to magnetic beads. Those skilled in the art will appreciate that other supports may be used in place of the magnetic beads, such as membranes, glass matrices or other types of beads, such as gold beads; methods of immobilization to these surfaces are known in the art.

[0287] Magnetic beads (MB) have been widely used in many biological applications such as separation, preconcentration and assay 57,58 . The beads are easy to use and can be removed from the sample using a magnetic rack without the need for centrifugation, sedimentation or filtration procedures. For sensors that can be used in the field and at home without a laboratory-based setup, MB can be used.

[0288] Streptavidin-coated MB was used because of its high efficiency for immobilization of biotin-modified DNA 59...

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Abstract

A general methodology for the development of highly sensitive and selective sensors that can achieve portable, low-cost and quantitative detection of a broad range of targets using only a personal glucose meter (PGM) is disclosed. The method uses recognition molecules that are specific for a target agent, enzymes that can convert an enzyme substrate into glucose, and PGM. Also provided are sensors, which can include a solid support to which is attached a recognition molecule that permits detection of a target agent, wherein the recognition molecule specifically binds to the target agent in the presence of the target agent but not significantly to other agents as well as an enzyme that can catalyze the conversion of a substance into glucose, wherein the enzyme is attached directly or indirectly to the recognition molecule, and wherein in the presence of the target agent the enzyme can convert the substance into glucose. The disclosed sensors can be part of a lateral flow device. Methods of using such sensors for detecting target agents are also provided.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Application No. 61 / 348,615, filed May 26, 2010, which is incorporated herein by reference in its entirety. technical field [0003] The present application relates to sensors, kits including the sensors, and methods of making and using the sensors. The sensor allows detection of a wide range of target agents, such as nucleic acids (eg DNA and RNA), proteins, toxins, pathogens, cells and metals, and can be used in combination with ready-to-use personal glucose meters. [0004] Government funding [0005] This invention was made with government support under DE-FG02-08ER64568 awarded by the US Department of Energy, ES16865 awarded by the National Institutes of Health, and CTS-0120978 awarded by the National Science Foundation. The US Government has certain rights in this invention. Background technique [0006] The development of portable and inexpensive sensors for q...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/00
CPCC12Q1/54C12Q1/34G01N33/558
Inventor 陆艺向宇
Owner THE BOARD OF TRUSTEES OF THE UNIV OF ILLINOIS
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