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Digital lspr for enhanced assay sensitivity

a digital lspr and assay technology, applied in the field of enhanced assay sensitivity, can solve the problems of insufficient detection of analytes in samples in minute quantities, and requires extremely sensitive assays for detection

Inactive Publication Date: 2015-09-03
LAMDAGEN CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes a method and system for detecting and measuring the concentration of analytes in a sample. The method involves capturing images of a sensor surface that can sustain a localized surface plasmon resonance (SPR) and analyzing changes in color over time. The system includes an optical imaging device and a processor for selecting areas of the sensor surface and measuring the change in color. The method has a better limit of detection for analytes than previous methods and can detect as little as 1 ng / mL. The system can be used with various types of analytes and can be integrated into a healthcare decision-making process.

Problems solved by technology

Often, molecules may be present in samples of interest only at very low concentrations and may require extremely sensitive assays for detection.
While a variety of assay procedures and detection techniques exist, they are often insufficient to detect analytes that are present in samples in minute quantities.

Method used

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  • Digital lspr for enhanced assay sensitivity
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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0086]Red Channel changes in response to a bulk refractive index change

[0087]LSPR sensors were imaged using a ThorLabs CMOS camera (DCC1645C-HQ) with a 10× magnification objective placed ˜7 mm from the surface (and ˜3-4 mm from the top of a flow cell cover). In the flow cell, different solutions were pumped successively (water, 5% EG, 10% EG, and 20% EG). For each solution, a series of images were acquired and analyzed. For each series of images, pixels were selected at random and 3×3 pixels centered on the randomly selected pixel were analyzed for changes in red channel values. The sensor surface raw data collected from a color CMOS sensor shows how the red channel changes for 3×3 pixels subsection of the entire image when the bulk refractive index is successively changed from water to 5% Ethylene Glycol (EG), 10% EG, and 20% EG (shown in FIG. 11). Panel A shows a raw signal for one randomly selected pixel, while panel B shows the same signal run through an averaging filter to remo...

example 2

Limit of Detection Determination Based on Detection of Analytes at Various Surface Coverage

[0090]FIG. 9 illustrates the detection of analytes at several surface coverages using digital LSPR, in accordance with embodiments. In a proof of concept experiment, a surface was prepared by co-adsorbing biotinylated IgG and native IgG at various ratios, but at a fixed total antibody concentration of 1 mg / mL (biotinylated IgG plus native IgG). Ratios of 10−6, 3*10−6, 8*10−6, 10−5, 10−4, and 10−3 were specifically prepared. The total concentration was large enough to saturate the surface. The surface coverage in biotinylated IgG was assumed to correspond to the ratio of biotinylated IgG to total antibody. Hence a surface coverage of 10−6 meant that during co-adsorption of biotinylated IgG and native IgG there were 1 biotinylated IgG per 106 native IgG. These model surfaces mimic the coverage of antigen in real assays. The biotinylated antibodies on the LSPR surface was detected using streptavi...

example 3

Prophetic

Detection of Nucleic Acid Target Molecules

[0092]In some embodiments, the disclosed digital LSPR methods and systems may be used for detection of target nucleic acid sequences in biological or environmental samples. One of skill in the art can envision immobilizing capture probes on the LSPR sensor surface that are complementary to a portion of one or more target nucleic acid sequences. Immobilization of probes on the surface may be accomplished by any of a variety of techniques known to those of skill in the art, for example by using patterned, self-assembled monolayers of thiolated oligonucleotides (or suitable linker molecules, as necessary) on noble metal surfaces. The surface may then be contacted with a sample of interest under conditions that promote hybridization of complementary sequences such that target nucleic acid molecules, if present, are captured and immobilized on the sensor surface. The sensor surface may then be rinsed and contacted with a solution compris...

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Abstract

Systems, methods, and devices related to detecting a presence of an analyte and / or determining a concentration of analytes are provided. An analyte may be provided on an LSPR-active surface. The LSPR-active surface may comprise sensitivity enhancing labels. The analyte may induce a local change near the LSPR-active surface. The LSPR-active surface may be imaged with an imaging device for images before, during, or after a reaction takes place. Local regions of interest within the images may be analyzed to detect the local changes.

Description

CROSS-REFERENCE[0001]This application claims the benefit of U.S. Provisional Application No. 61 / 966,576, filed Feb. 26, 2014, and U.S. Provisional Application No. 62 / 108,979, filed Jan. 28, 2015, which applications are incorporated herein by reference.BACKGROUND OF THE INVENTION[0002]The accuracy, sensitivity, reproducibility, and ease-of-use of instruments designed for detection and quantitation of specific molecules (e.g., analytes or markers) and / or analysis of molecular interactions are of paramount concern in a variety of fields including biomedical research, clinical diagnostics, environmental testing, and industrial process monitoring. These concerns are driven by a variety of factors including the difficulty and cost associated with producing and / or isolating molecules of interest in biomedical research, for example, or the critical impact that a diagnostic test result may have on proper diagnosis and treatment of disease in the healthcare field. Often, molecules may be pres...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N21/65C12Q1/68
CPCG01N33/54366G01N33/54373G01N2201/125G01N21/658C12Q1/6837G01N21/554C12Q1/6825C12Q2565/628
Inventor GERION, DANIELESTORER, RANDOLPH
Owner LAMDAGEN CORP
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