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Fluorescent magnetic bead micro-fluidic chip and analytical instrument thereof

A microfluidic chip and magnetic bead technology, which is applied in fluid controllers, instruments, and material analysis through optical means, can solve the problems of complex reaction process, many reagent components, and poor reaction effect, and achieve simple operation, The effect of less reagent consumption and less sample volume

Active Publication Date: 2021-01-19
广州华澳生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Existing microfluidic chips are mainly driven by centrifugation and capillary action, and the mixing and reaction adopts a curved pipe or a built-in column in the mixing zone, which has low mixing efficiency and poor reaction effect. For example, the publication numbers are "CN106807461A" and "CN108181458A" Chinese patent; In addition, the microfluidic chip used for quantitative detection uses chemiluminescence method, the reagent components are many, and the reaction process is complicated, resulting in a multi-layer structure of the microfluidic chip, and the preparation is cumbersome, such as the Chinese patent with the publication number "CN201510696706"

Method used

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  • Fluorescent magnetic bead micro-fluidic chip and analytical instrument thereof
  • Fluorescent magnetic bead micro-fluidic chip and analytical instrument thereof
  • Fluorescent magnetic bead micro-fluidic chip and analytical instrument thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Below, with figure 1 The chip structure of the negative pressure one-step method is applied to the detection of C-reactive protein (CRP) as an example (double-antibody sandwich method detection), and the present invention is described:

[0041] (1) Antibody labeling

[0042] Add 1 mg of magnetic beads (polystyrene-coated iron compound, 3 μm in diameter, surface-modified carboxyl group), 10 μg of EDC and 15 μg of NHS solution and 20 μg of anti-CRP monoclonal antibody (CRP primary antibody) solution to 1 ml of 10 mM pH7.5 phosphate buffer solution, mix Uniform and react at room temperature for 4h, add 1mg lysine to block. Enrich with a magnet, remove unreacted antibodies, add 0.1% BSA 0.01M pH7.5 PBS solution 1ml to redissolve, and obtain CRP primary antibody-labeled capture magnetic beads solution.

[0043] Take 1ml of fluorescent microspheres with a solid content of 1% (europium compound coated with polystyrene, diameter 200nm, surface modified carboxyl group), add 15...

Embodiment 2

[0053] Below, with figure 2 The chip structure of the two-step negative pressure method is applied to the detection of serum amyloid A (SAA) as an example to illustrate the present invention:

[0054] (1) Antibody labeling

[0055] Add 1 mg of magnetic beads (polystyrene-coated iron compound, 3 μm in diameter, surface-modified carboxyl group), 10 μg of EDC and 15 μg of NHS solution and 15 μg of anti-SAA monoclonal antibody (SAA primary antibody) solution to 1 ml of 10 mM pH7.5 phosphate buffer solution, mix Uniform and react at room temperature for 4h, add 1mg lysine to block. Enrich with a magnet, remove unreacted components, add 0.1% BSA 0.01M pH7.5 PBS solution 1ml to redissolve, and obtain the SAA primary antibody-labeled capture magnetic bead solution.

[0056] Take 1ml of fluorescent microspheres with a solid content of 1% (europium compound coated with polystyrene, 200nm in diameter, carboxyl group modified on the surface), add 15mg of EDC and 50mg of sulfo-NHS solut...

Embodiment 3

[0065] Below, with image 3 The chip structure of the positive pressure one-step method is applied to the detection of 25 hydroxyvitamin D as an example (competitive method detection), and the present invention is described:

[0066] (1) Antibody labeling

[0067] Add 1 mg of magnetic beads (polystyrene-coated iron compound, 3 μm in diameter, surface-modified carboxyl group), 10 μg of EDC and 15 μg of NHS solution and 20 μg of BSA-coupled 25-hydroxyvitamin D (VD-BSA) into 1 ml of 10 mM pH7.5 phosphate buffer solution, mixed evenly and reacted at room temperature for 4h, adding 1mg of lysine to block. Enrich with a magnet, remove unreacted components, add 0.1% BSA 0.01M pH7.5 PBS solution 1ml to redissolve, and obtain VD-BSA labeled capture magnetic bead solution.

[0068] Take 1ml of fluorescent microspheres with a solid content of 1% (polystyrene coated europium compound, 200nm in diameter, surface modified carboxyl group), add 15mg EDC and 50mg sulfo-NHS solution and 30μg 25...

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Abstract

The invention discloses a fluorescent magnetic bead micro-fluidic chip which can be divided into four different layouts according to driving force and reaction steps, namely a negative-pressure one-step method, a negative-pressure two-step method, a positive-pressure one-step method and a positive-pressure two-step method. The invention also discloses an analytical instrument with the fluorescentmagnetic bead micro-fluidic chip, wherein the analytical instrument comprises an instrument frame, a quantitative sample adding device, a cleaning liquid storage bin, a waste liquid storage bin, a negative pressure control pump, an air pump extrusion device, a reaction zone magnetic field, a luminescence detection system, a control analysis module, a software system and the like. After the chip isplaced in the instrument, clicking is carried out to start a test, and the instrument can automatically complete all operations.

Description

technical field [0001] The invention relates to a highly sensitive and quantitative detection system for analytes using fluorescent microspheres, magnetic beads, microfluidic chips and analytical instruments, which can detect pathogens, major diseases (such as tumors, cardiovascular diseases), illegal drugs, and drugs Accurate, highly sensitive, quantitative detection of analytes such as , food safety, etc., belongs to the field of microfluidic chip detection technology. Background technique [0002] Microfluidic core technology is to integrate the basic operation units of sample preparation, reaction, separation and detection in the process of biological, chemical and medical analysis into a chip through micro-fabrication technology, which has the advantages of miniaturization, integration, fast analysis speed, reagent Significant advantages such as less consumption. [0003] Existing microfluidic chips are mainly driven by centrifugation and capillary action, and the mixi...

Claims

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Application Information

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IPC IPC(8): B01L3/00G01N21/64G01N33/543G01N35/00
CPCB01L3/50273B01L3/502761G01N35/00G01N33/54326G01N21/6428G01N2035/00099G01N2035/00237G01N2021/6439B01L2200/0668B01L2200/10B01L2400/043B01L2300/0654
Inventor 于龙波于永涛黎权
Owner 广州华澳生物科技有限公司
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