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Confining liquid for liquid chip, confining method and application

A liquid chip, sealing liquid technology, applied in material inspection products, measuring devices, instruments, etc., can solve the problems of high reaction background, influence of reaction precision and accuracy and accuracy, influence of reaction sensitivity, etc. Sensitivity and precision, reduced non-specific binding, reduced background effects

Pending Publication Date: 2020-02-25
北京协和洛克生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, in the traditional magnetic bead coating method, after the microspheres are activated by NHS and EDC, the microspheres bind and couple with the protein through their own carboxyl groups to obtain magnetic beads coated with antibodies. The coated microspheres are directly Stored in phosphate buffer containing 1% BSA, it plays the role of sealing and preservation, and the sealing of the coated magnetic beads is not carried out separately, so that the gaps on the microspheres cannot be completely and fully closed during the reaction process. The background of the reaction will be higher, which will affect the sensitivity of the reaction, and also have a certain impact on the precision and accuracy of the reaction.

Method used

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  • Confining liquid for liquid chip, confining method and application
  • Confining liquid for liquid chip, confining method and application
  • Confining liquid for liquid chip, confining method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] This example is used to illustrate the preparation of phosphate buffer.

[0062] Prepare phosphate buffer solution, accurately weigh out 0.358g Na2HPO4·12H2O, 0.024g KH2PO4, 0.8g NaCl, 0.02g KCl reagent with an electronic balance and place it in a 100ml beaker, add 80% of the purified water, stir well, and adjust the pH to 7.4. Then set the volume for use.

Embodiment 2

[0064] This example is used to illustrate the preparation of the sealing liquid of the present invention.

[0065] Prepare a blocking solution to improve the performance of coated antibody magnetic beads. Weigh 3g bovine serum albumin and 2g glycine. Use a pipette to measure 0.05ml Tween 20 and 0.1ml proclin 300. Add the reagent to 80ml phosphate buffer. After completely dissolved, use a volumetric flask to make the volume up to 100ml with the prepared phosphate buffer.

Embodiment 3

[0067] This example is used to illustrate the sealing method of the present invention, taking the sealing liquid in Example 2 as an example.

[0068] The antibody coating process of magnetic beads:

[0069] 3.1 Experimental materials:

[0070] Magnetic beads: purchased at Luminex 1.25X10 7 / ml,1mL.

[0071] Activation reagent: 100 mM sodium dihydrogen phosphate pH 6.3.

[0072] Reagent: 50mM MES, pH5.0.

[0073] Magnetic beads storage solution: 10mM phosphate buffer, pH7.4, 1% BSA, 0.05% Tween 20.

[0074] 3.2 Coating process:

[0075] 3.2.1 Wash the microspheres

[0076] (1) Resuspend the magnetic microspheres, oscillate at 2000 rpm for 15 minutes, and transfer 5.0x 10 6 Magnetic microspheres into the centrifuge tube.

[0077] (2) Put the centrifuge tube containing magnetic beads into the magnetic separator and let stand for 30-60 seconds, remove the supernatant, and be careful not to touch the magnetic beads.

[0078] (3) Remove the tube from the magnetic separator, add 200μL of purified wa...

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PUM

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Abstract

The invention provides confining liquid for a liquid chip. The confining liquid comprises a phosphate buffer solution, protein, amino acid, a surfactant and a preservative. According to the confiningliquid, a confining link of coating antibody magnetic beads is added in the traditional liquid chip magnetic bead coating process, and the amino acid and the protein in the confining liquid can be well combined with gaps in the magnetic beads, so that the nonspecific binding of serum protein is reduced, the background of the reaction is reduced, and the sensitivity and precision of the reaction are improved.

Description

Technical field [0001] The invention relates to the technical field of biological detection reagents, and more specifically, to a sealing solution, a sealing method and an application for liquid chip technology. Background technique [0002] Liquid chip technology is also called "liquid suspension chip technology", "multifunctional flow lattice technology", "liquid chip analysis technology", "flow fluorescence detection technology", "multifunctional multi-index parallel analysis technology" in China. , "(Microsphere) Suspended Array Technology" etc. It organically integrates a number of scientific and technological achievements such as fluorescent coded microsphere technology, laser analysis technology, flow cell technology, high-speed digital signal processing technology, computer algorithms, etc., with free combination, high throughput, high speed, low cost, and accuracy High, good repeatability, high sensitivity, wide linear range, no washing, easy operation, protein and nucl...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/543
CPCG01N33/5306G01N33/54326
Inventor 李捷吴建榕马奕晶张丹浩张金红
Owner 北京协和洛克生物技术有限责任公司
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