Biological macromolecule detection method based on nano homogeneous time-resolved fluoroimmunoassay and droplet-based micro-fluidic technology

Abstract

The invention belongs to the field of micro-fluidic chips, and relates to a biological macromolecule detection method. An alpha-LISA technology and a droplet-based microfluidics technology are combined and optimized to prepare a micro-fluidic chip liquid. A novel biological macromolecule detection method with a high sensitivity is provided. A three-phase mixing technology is adopted. A passive mixing structure is used to fully and evenly mix the reaction liquid. A cross focusing technology is used to form droplets. The generated droplets have the advantage of smaller volume, and thus the reactions become faster. Moreover, the requirements on the samples are lower, the reactions are specific, sensitive, fast, and full; the washing does not need to be carried out after reactions; complicated complexes such as complete proteins, enzyme complex, bacteriophage, and the like, can be detected; the technical bottleneck of conventional commercial liquid chips at present can be broken through; the provided method can be applied to fast clinical biological macromolecule detection; the detection sensitivity and specificity are both improved, and the required sample amount is reduced.

Description

technical field

[0001] The invention belongs to the field of biomedical research and clinical application, and relates to a detection method for biological macromolecules; in particular, it relates to a microfluidic chip based on nano-homogeneous time-resolved fluorescence immunity and droplet microfluidic technology and a method for detecting biological macromolecules . Background technique

[0002] The prior art discloses that the liquid phase chip technology is a modern biochip technology developed under the background of post-genomics with functional genomics and proteomics as the research objects. This technology utilizes the high specific recognition ability and high-strength binding ability of antigen-antibody to capture specific targets for further qualitative and quantitative research, such as detection of protein markers, capture of tumor cells, etc. Compared with the traditional solid-phase chip based on the principle of ELISA, the liquid-phase chip has the advan...

Examples

Embodiment 1

[0066] combine as figure 1 , figure 2 The Alpha-LISA technology and droplet-based microfluidics technology are shown to prepare anti-human carcino-embryonic antigen (carcino-embryonic antigen, CEA, commercially available) droplet microfluidic chip:

[0067] 1. Design of microfluidic chip, such as figure 1 Shown includes the following parts:

[0068] ①The three-way liquid-phase inflow pipeline of the sample to be tested (standard product), donor microspheres and their buffer, and acceptor microspheres ( figure 1 Middle A, figure 1 Middle B, figure 1 Middle C);

[0069] ②The fishbone-shaped passive hybrid structure is connected at the end of the confluence of the three-way pipes ( figure 1 middle D, figure 2 );

[0070] ③ Both sides of the end of the passive mixing structure are vertically connected to the oil phase inflow channel, and a droplet microreactor will be formed at the junction of the water and oil phases during the reaction ( figure 1 Middle E);

...

technical field

[0001] The invention belongs to the field of biomedical research and clinical application, and relates to a detection method for biological macromolecules; in particular, it relates to a microfluidic chip based on nano-homogeneous time-resolved fluorescence immunity and droplet microfluidic technology and a method for detecting biological macromolecules . Background technique

[0002] The prior art discloses that the liquid phase chip technology is a modern biochip technology developed under the background of post-genomics with functional genomics and proteomics as the research objects. This technology utilizes the high specific recognition ability and high-strength binding ability of antigen-antibody to capture specific targets for further qualitative and quantitative research, such as detection of protein markers, capture of tumor cells, etc. Compared with the traditional solid-phase chip based on the principle of ELISA, the liquid-phase chip has the advan...

Examples

Embodiment 1

[0066] combine as figure 1 , figure 2 The Alpha-LISA technology and droplet-based microfluidics technology are shown to prepare anti-human carcino-embryonic antigen (carcino-embryonic antigen, CEA, commercially available) droplet microfluidic chip:

[0067] 1. Design of microfluidic chip, such as figure 1 Shown includes the following parts:

[0068] ①The three-way liquid-phase inflow pipeline of the sample to be tested (standard product), donor microspheres and their buffer, and acceptor microspheres ( figure 1 Middle A, figure 1 Middle B, figure 1 Middle C);

[0069] ②The fishbone-shaped passive hybrid structure is connected at the end of the confluence of the three-way pipes ( figure 1 middle D, figure 2 );

[0070] ③ Both sides of the end of the passive mixing structure are vertically connected to the oil phase inflow channel, and a droplet microreactor will be formed at the junction of the water and oil phases during the reaction ( figure 1 Middle E);

...

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