Integration micro-fluidic chip for immune analysis research and applications thereof

A microfluidic chip and immune analysis technology, applied in the field of immune analysis, can solve the problems of long analysis time and cumbersome steps, and achieve the effects of improving analysis efficiency, shortening reaction time, and reducing consumption

Inactive Publication Date: 2013-04-10
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The steps of traditional ELISA are cumbersome and mostly manual, requiring a large amount of expensive immunological reagents, and the analysis time is long, usually several hours

Method used

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  • Integration micro-fluidic chip for immune analysis research and applications thereof
  • Integration micro-fluidic chip for immune analysis research and applications thereof
  • Integration micro-fluidic chip for immune analysis research and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The microfluidic chip is used for the research application of the optimal concentration of drugs. image 3 After surface modification of the chip shown with BSA-glutaraldehyde-Protein A, 5 μg / The antibody in ml was passed into the PDMS channel and allowed to stand for 60 minutes, and then passed into TBS buffer to wash away the unfixed antibody. Pass 5 μg / ml antigen into the channel and let it stand for 20 minutes, then pass through TBS buffer to wash away unfixed antigen. Pass 5 μg / ml of secondary antibody into the channel and let it stand for 20 minutes, then pass through TBS buffer to wash away unfixed secondary antibody. Pass through 4-MUP with a drug concentration of 0.0001 mmol / l to react for 3 minutes, and then place it in a microplate reader to detect the fluorescence intensity (excitation wavelength and emission wavelength are 365nm and 448nm, respectively). Repeat the above operation, respectively change the concentration of the drug 4-MUP to 0.001, 0.01, 0...

Embodiment 2

[0030] The application of microfluidic chips in the study of drug action time. image 3 After surface modification of the chip shown with BSA-glutaraldehyde-protein A, 5 μg The / ml antibody was passed into the PDMS channel and allowed to stand for 60 minutes, and then passed into TBS buffer to wash away the unfixed antibody. Pass 5 μg / ml antigen into the channel and let it stand for 20 minutes, then pass through TBS buffer to wash away unfixed antigen. Pass 5 μg / ml of secondary antibody into the channel and let it stand for 20 minutes, then pass through TBS buffer to wash away unfixed secondary antibody. After passing through 4-MUP with a drug concentration of 0.10mmol / l, place it under a microplate reader to detect the fluorescence intensity (excitation wavelength and emission wavelength are 365nm and 448nm respectively). The experimental results are as follows: Figure 5 shown.

Embodiment 3

[0032] Application of microfluidic chip in the study of inhibition rate of enzyme inhibitors. image 3 After surface modification of the chip shown with BSA-glutaraldehyde-protein A, 5 μg The / ml antibody was passed into the PDMS channel and allowed to stand for 60 minutes, and then passed into TBS buffer to wash away the unfixed antibody. Pass 5 μg / ml antigen into the channel and let it stand for 20 minutes, then pass through TBS buffer to wash away unfixed antigen. Pass 5 μg / ml of secondary antibody into the channel and let it stand for 20 minutes, then pass through TBS buffer to wash away unfixed secondary antibody. One inlet of the chip is fed with 0.10mM 4-MUP and inhibitor KH 2 PO 4 buffer, and the other inlet into 0.10mM 4-MUP and 1mM KH 2 PO 4 React for 3 minutes, and then place it under a microplate reader to detect the fluorescence intensity (excitation wavelength and emission wavelength are 365nm and 448nm, respectively). Experimental results such as Figure ...

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Abstract

The invention discloses an integration micro-fluidic chip for immune analysis research. The chip is mainly composed of a concentration gradient generation unit, an equal allocation unit and a micropore reaction chamber matrix with the diameter of 5.8mm and the depth of 100 mu, and has small volume and large specific surface area; the arrangement of the micropore reaction chamber matrix is consistent with the arrangement of a traditional porous plate, and the micropore matrix can be applied to a standard microplate reader for detecting and gathering analysis data. According to the invention, the operations of absorption and fixation of antibodies, antigens and enzyme during immune analysis operation, the preparation of sample solutions with different concentrations, liquid replacement, washing and the other operations are all integrated on a chip and are completed automatically according to the process creatively; the reagent consumption can be lowered obviously; the analysis time can be shortened; and the efficiency can be improved. Compared with the traditional porous plate technology, according to the invention, the complex manual operations of sample solution preparation, washing and the like are omitted, and the integration micro-fluidic chip for immune analysis research is compatible with the standard microplate reader on the detecting method.

Description

technical field [0001] The invention relates to immune analysis technology, in particular to an integrated microfluidic chip for immune analysis research and its application. Background technique [0002] Immunoassay methods have extremely high selectivity and sensitivity, and can be widely used in clinical diagnosis, proteomics, drug research, biological research, environmental analysis and other fields. Conventional immunoassays are divided into homogeneous immunoassays and heterogeneous immunoassays, and enzyme-linked immunosorbent assay (ELISA) is the most common and widely used heterogeneous immunoassay, and they are all carried out in 96 microwell plates . The operation usually includes multiple operations such as coating antibodies and antigens, repeated washing, drying, blocking, labeling antibodies, and incubation reactions. The steps of traditional ELISA are cumbersome and mostly manual, requiring a large amount of expensive immunological reagents, and the analys...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N21/33G01N21/64B01L3/00
Inventor 成志毅侯凤华
Owner SUN YAT SEN UNIV
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