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Micro-scale magnetic concentration detection device and method for immunoassay

An immunoassay and detection device technology, which is applied in the direction of measuring devices, analysis materials, instruments, etc., can solve the problems of miniaturized immunoassay sensitivity that needs to be improved, and meet the production requirements and low cost, less sample consumption, and ensure stability. Effect

Inactive Publication Date: 2019-07-19
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the results of immunoassays on some microfluidic platforms are comparable to those of traditional methods, the sensitivity of most miniaturized immunoassays still needs to be improved

Method used

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  • Micro-scale magnetic concentration detection device and method for immunoassay
  • Micro-scale magnetic concentration detection device and method for immunoassay
  • Micro-scale magnetic concentration detection device and method for immunoassay

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Example 1: Evaluation of Signal Amplification Effect of Microscale Magnetic Enrichment

[0048] In this embodiment, magnetic beads labeled with horseradish peroxidase (HRP) are used as a simulated magnetic immune complex to evaluate the effect of the present invention. The specific detection method is as follows:

[0049] (1) Preparation of simulated magnetic immune complex (HRP-labeled magnetic beads): In a clean 10mL centrifuge tube, add 0.5mL core-shell magnetic microsphere suspension with different particle sizes (0.5μm, 2.5μm, 5.6μm) (5mg / mL) and 7mL 2.5% glutaraldehyde solution, mixed on a mixer for 30s, sealed, and shaken at room temperature for 3h; placed the centrifuge tube on a magnetic stand for magnetic separation, discarded the supernatant, and washed twice with ultrapure water Finally, add 5 mL of PBS buffer solution containing 10 mg / mL HRP, mix well, seal, and shake at room temperature for 12 hours to fully suspend the magnetic microspheres to ensure that...

Embodiment 2

[0052] Example 2: Detection of Carcinoembryonic Antigen in Serum

[0053] In this embodiment, carcinoembryonic antigen (CEA) in serum is used as the target object, and the microscale magnetic enrichment in situ detection method is as follows:

[0054] (1) Sample pretreatment: take a serum sample and properly dilute it with PBS; prepare a series of CEA standard solutions with different concentrations at the same time.

[0055] (2) Preparation of magnetic immune complex: In a clean 0.5mL sterilized centrifuge tube, add 50μL standard solution or diluted serum sample 32 and 50μL capture antibody-labeled magnetic beads 31, and incubate at room temperature at 820rpm for 2h; Place the centrifuge tube on a magnetic stand, discard the liquid after 1 min, add 100 μL of washing solution, discard the liquid again after standing for 1 min, and repeat this washing operation three times; add 50 μL of biotinylated detection antibody 34 to the centrifuge tube, Incubate at 820 rpm for 1 hour, ...

Embodiment 3

[0059] Example 3: Detection of Prostate Specific Antigen in Serum

[0060] In this embodiment, prostate-specific antigen (PSA) in serum is used as the target object, and the microscale magnetic enrichment in situ detection method is as follows:

[0061] (1) Sample pretreatment: take a serum sample and properly dilute it with PBS; prepare a series of PSA standard solutions with different concentrations at the same time.

[0062] (2) Preparation of magnetic immune complex: In a clean 0.5mL sterilized centrifuge tube, add 50μL standard solution or diluted serum sample and 50μL capture antibody-labeled magnetic beads, and incubate at room temperature at 820rpm for 2h; put the centrifuge tube Place on a magnetic stand, discard the liquid after 1 min, add 100 μL of washing solution, discard the liquid again after standing for 1 min, and repeat this washing operation three times; add 50 μL of biotinylated detection antibody to the centrifuge tube, and rotate at 820 rpm at room temper...

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Abstract

The invention relates to a micro-scale magnetic concentration detection device and a method for immunoassay. The micro-scale magnetic concentration detection device includes a micro-fluidic chip whichincludes one or more micro-channel and the micro-channels comprises a sample pool used for adding magnetic immune complexes and enzyme substrates, a detection zone and a sample outlet. The magnetic immune complexes are formed by biological samples connected with magnetic beads and enzymes through specific immune responses. The micro-scale magnetic concentration detection device further includes amagnet which is arranged above the detection zone. The invention further provides a detection method of using the micro-scale magnetic concentration detection device. According to the micro-scale magnetic concentration detection method, samples used for immunoassay are concentrated in the detection zone of the micro-fluidic channels, and by utilizing the features of rapid mass transfer of reporter molecules of micro-scale, the detection limit of targeted samples is decreased by more than two orders of magnitude.

Description

technical field [0001] The invention relates to the technical field of microfluidic immune analysis, in particular to a microscale magnetic enrichment detection device and detection method for immune analysis. Background technique [0002] Immunoassay is a biochemical test that uses antigen-antibody specific reactions to evaluate the presence or concentration of target molecules in a solution, and is also one of the most commonly used detection methods in clinical diagnosis and related research fields. Immunoassays, including enzyme-linked immunosorbent assay (ELISA), are widely used in the detection of biomarkers due to their rapidity, sensitivity, good selectivity, and high cost-effectiveness. However, many traditional immunoassays are still labor-intensive, with long detection times and high sample and solvent losses, which limit the scope of immunoassay applications. [0003] Miniaturized immunoassays implemented on microfluidic platforms can overcome some of the limita...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543
CPCG01N33/535G01N33/54326
Inventor 李攻科董建伟杨佳妮夏凌
Owner SUN YAT SEN UNIV
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