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Improved proteomic multiplex assays

A technology for testing samples, diluents, applied in the field of proteomics assays, which can solve the problems of limitations, false detection signals, etc.

Pending Publication Date: 2021-04-09
私募蛋白质体运营有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the sensitivity and specificity of many assay formats is limited by the ability of detection methods to resolve true signals from signals due to non-specific associations during the assay and generate false detection signals

Method used

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  • Improved proteomic multiplex assays
  • Improved proteomic multiplex assays
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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0222] Preparation of oligonucleotides

[0223] The automated synthesis of oligodeoxynucleosides is a routine practice in many laboratories (see for example Matteucci, M.D. and Caruthers, M.H., (1990) J.Am.Chem.Soc., 103 :3185-3191, the contents of which are hereby incorporated by reference in their entirety). The synthesis of oligoribonucleosides is also well known (see for example Scaringe, S.A., et al., (1990) Nucleic Acids Res. 18 :5433-5441, the contents of which are hereby incorporated by reference in their entirety). As described herein, phosphoramidites can be used to incorporate modified nucleosides into oligonucleotides by chemical synthesis, and triphosphates can be used to incorporate modified nucleosides into oligonucleotides by enzymatic synthesis. (See eg, Vaught, J.D. et al. (2004) J.Am.Chem.Soc., 126 : 11231-11237; Vaught, J.V., et al. (2010) J.Am.Chem.Soc. 132 , 4141-4151; Gait, M.J. "Oligonucleotide Synthesis a practical approach" (1984) IRL Press (Oxfor...

Embodiment approach

[0236] In some embodiments, a method is disclosed comprising: a) contacting a first test sample with a first set of aptamers to form a first mixture, wherein the first test sample is Z% of a biological sample Dilution, where Z is 5% to 39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16% of the biological sample %, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, or 39%) dilutions, and at least A is present in the first set of aptamers 3 different aptamers; b) contacting a second test sample with a second set of aptamers to form a second mixture, wherein said second test sample is a Y% dilution of said biological sample, wherein Y is less than Z, and wherein at least A is present in the second set of aptamers 2 c) contacting a third test sample with a third set of aptamers to form a third mixture, wherein said third test sample is an X% dilution of said biological sample, where X is less than Y, and In sa...

Embodiment 1

[0378] Example 1. Multiple aptamer analysis of samples

[0379] This example describes a multiplex aptamer assay for the analysis of samples and controls.

[0380] Multiplex aptamer assay method

[0381] All steps of the multiplex aptamer assay were performed at room temperature unless otherwise indicated.

[0382] Preparation of aptamer master mix solution.

[0383] The 5272 aptamers were grouped into three unique mixtures, Dil1, Dil2, and Dil3, and corresponded to 20%, 0.5%, and 0.005% plasma or serum sample dilutions, respectively. Assignment of the aptamers to the mixture was determined empirically by assaying a dilution series of matched plasma and serum samples with each aptamer and identifying the sample dilutions that gave the most linear signal range. Isolation of the aptamer and mixing with different dilutions (20%, 0.5% or 0.005%) of plasma or serum samples allowed the assay to span 10 7 fold range of protein concentration. A stock solution of the aptamer maste...

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Abstract

Methods, devices, reagents and kits designed to improve the performance of proteomic based assays are provided. Such methods have a wide utility in proteomic applications for research and development, diagnostics and therapeutics by providing for a reduction or elimination of background signal and improved specificity for protein binding reagents in a multiplex assay formats.

Description

technical field [0001] The present disclosure relates generally to the field of proteomic assays, and methods, devices, reagents and kits designed to improve the performance of such assays. Such methods have broad utility in proteomics applications for research and development, diagnostics, and therapeutics. In particular, materials and methods are provided for reducing or eliminating background signal and increasing the specificity of protein binding reagents in multiplex assay formats. Background technique [0002] Assays for the detection and quantification of physiologically important molecules in biological samples and other sample types are important tools in the fields of scientific research and healthcare. For example, multiplex array assays employ surface-bound probes to detect target molecules in a sample. Surface bound probes may be oligonucleotides, peptides, polypeptides, proteins, antibodies, affibodies, aptamers or other molecules (collectively referred to a...

Claims

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Application Information

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IPC IPC(8): C07B61/00C07H19/00C07H19/073G01N33/53
CPCG01N33/6803G01N33/5306G01N33/5308G01N2570/00C12Q1/6848
Inventor S·克雷默E·卡蒂留斯D·济奇
Owner 私募蛋白质体运营有限公司
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