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System and method for protein micromolecule enriching-detection based on microfluidic chip

A small molecule, enrichment technology, applied in the field of biomedical detection, can solve the problems of difficult to achieve fast and simple detection, difficult to detect cytokines, insufficient sensitivity, etc., to reduce the cost of enrichment and detection, and save space resources And the effect of saving power resources and human resources

Active Publication Date: 2017-08-18
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the detection methods of small protein molecules such as cytokines mainly include ELISA, immunofluorescence detection, immunoelectrochemical detection, surface plasmon resonance, etc., all of which involve antibody incubation, sample incubation, labeling, and many elution steps. It is cumbersome, it is difficult to achieve fast and simple detection, and it is difficult to perform detection under the conditions of limited space and equipment
[0004] However, the detection limit of the current immunochromatographic test strips is usually tens of ng / mL, the sensitivity is insufficient, and it is difficult to detect low-level cytokines

Method used

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  • System and method for protein micromolecule enriching-detection based on microfluidic chip
  • System and method for protein micromolecule enriching-detection based on microfluidic chip
  • System and method for protein micromolecule enriching-detection based on microfluidic chip

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Embodiment 1

[0042] The present invention provides a microfluidic chip-based enrichment-detection system for small protein molecules, including an enrichment unit, a detection unit and a liquid circuit unit, such as figure 2 As shown, the enrichment unit coats the conjugate of nanoparticles and target small molecule protein monoclonal antibody on the enrichment chip 4, and the target small molecule protein in the detection solution specifically binds to the monoclonal antibody, and is enriched in The enrichment chip surface.

[0043] The detection unit is provided with a detection line and a quality control line successively on the detection chip 11 according to the flow direction of the detection liquid, and the detection line is coated with a polyclonal antibody of the target small molecular protein. After the antibody is combined, the detection line develops color, and the target small molecular protein is detected; the quality control line is coated with a secondary antibody that spec...

Embodiment 2

[0052] In order to realize the flow of nanoparticles from the enrichment unit to the detection unit, this embodiment provides an example, such as figure 1 , 3 As shown, the enrichment unit is composed of an enrichment chip and a conjugate of nanoparticles and monoclonal antibodies. The nanoparticles are superparamagnetic nanoparticles with a diameter of 100-300nm, and the diameter of the superparamagnetic nanoparticles is 100-300nm. The addition density on the enrichment chip is 12-20ug / mm3, and the thickness is 0.3-0.7mm.

[0053] Conjugates of superparamagnetic nanoparticles and monoclonal antibodies of target small molecular proteins are coated on the surface of the enrichment chip, and the enrichment chip is sequentially provided with a bottom plate 20 of the enrichment chip, a lower connection layer 19, and a middle channel layer from bottom to top. 18. The upper connection layer 17 and the upper cover plate 16, the enrichment chip surface is also provided with enrichmen...

Embodiment 3

[0057] Methods for enrichment and detection of IL-6.

[0058] Step 1: preparing a superparamagnetic nanoparticle-IL-6 antibody conjugate, which is used for the enrichment of the target cytokine interleukin 6-(IL-6). The specific preparation method is as follows:

[0059] a) Select commercially available superparamagnetic nanoparticles with a diameter of 100-300nm, take 1mg of magnetic particles into a 1.5ml centrifuge tube, wash 3 times with 500ul MEST (pH 6.0, 0.05% Tween 20), remove the upper surface after magnetic separation clear; add newly prepared 100ul EDC (5mg / ml) and 100ul NHS (5mg / ml) solutions to the centrifuge tube containing magnetic particles, and add 300ul PBST solution; activate at 4°C for 8h or at 25°C for 30-60min or 37 Activate at ℃ for 4 to 5 hours, during which the magnetic particles are kept in suspension; the centrifuge tube is placed on a magnetic separation rack for magnetic separation, the supernatant is removed, 500ul MEST is added, the magnetic par...

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Abstract

The invention relates to a system for protein micromolecule enriching-detection based on a microfluidic chip. The system comprises an enriching unit, a detection unit and a liquid path unit, wherein target micromolecule protein in detection liquid of the enriching unit is specifically combined with a monoclonal antibody; the detection unit is successively provided with a detection line and a quality control line on a detection chip in the flowing direction of the detection liquid, the detection line displays color, and the target micromolecule protein is detected out; the quality control line displays colors to verify whether the detection process is successful or not; a liquid path system is used for selectively controlling the detection liquid to pass through the enriching unit, the detection unit and a waste liquid tank. The system has the advantages that the enriching and detection processes are integrated on the basis of a chip technique, and the efficiency and accuracy are realized; the sensitivity for detecting the target micromolecule protein of the system is better than the sensitivity of the traditional test paper detection.

Description

technical field [0001] The invention belongs to the field of biomedical detection, and in particular relates to a microfluidic chip-based enrichment-detection system and method for small protein molecules. Background technique [0002] Microfluidic chip technology is a method that integrates sample preparation, enrichment, reaction, separation, and detection units in the process of biological, chemical, and medical analysis into the chip through the control of fluid at the micro scale, so as to achieve the purpose of automatic analysis. Technical means have become one of the most cutting-edge technologies in the world today, with the characteristics of miniaturization, miniaturization, controllable liquid flow, less reagent consumption, fast analysis speed, and easy integration and scale. Immunochromatography test strips are a fast, convenient and visual detection method. Using microfluidic chip technology combined with immunochromatography test strips to realize portable an...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 邓玉林李永瑞姚梦迪李昱樊云龙吕雪飞李晓琼王品虹
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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