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Chemiluminescent immunological detection system with microbubble-accelerated immunoreaction

A technology of chemiluminescence immunoassay and detection system, which is applied in the detection system of single-component chemiluminescence immunoassay, and the field of rapid immunoassay in which microbubbles accelerate the immune response. , improved reproducibility, and reduced analysis costs

Inactive Publication Date: 2009-09-30
JIANGSU CANCER HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Different methods have been used to accelerate the mass transfer rate, such as electric and magnetic field driving, trapped matrix rotation, and low-energy microwaves, etc., but these methods require complex or special devices

Method used

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  • Chemiluminescent immunological detection system with microbubble-accelerated immunoreaction
  • Chemiluminescent immunological detection system with microbubble-accelerated immunoreaction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Preparation of sensors based on antibody-modified resin microbeads.

[0034] The specific preparation process of the sensor is as follows: 50 mg of resin microbeads are swelled in 20% ethanol solution, rinsed three times with the activation buffer solution 2-(N-morpholine) ethanesulfonic acid of pH 5.5, and redispersed in the activation buffer solution. Then add 30 mg carbodiimide (EDC) and 30 mg N-hydroxysuccinimide (NHS), stir and activate at room temperature for 2 hours, wash with coupling buffer (pH7.0 phosphate buffer) three times, Add 200 μl of monoclonal antibody, stir slowly at room temperature for 2 hours, place in refrigerator at 4°C for 24 hours, then block with 1% bovine serum albumin for 24 hours, and wash thoroughly with coupling buffer. Finally, the antibody-modified resin microbeads were filled into a 4 cm long colorless transparent glass tube (with an inner diameter of 1.6 mm) to prepare the immunosensor.

Embodiment 2

[0035]Embodiment 2: in combination with figure 1 Flow Injection Chemiluminescent Immunoassay System for Accelerated Immune Response to Microbubbles is further illustrated:

[0036] The microbubble-accelerated flow injection chemiluminescence immunoassay system includes a solution delivery system, a microbubble-accelerated incubation device, an immunosensor, a chemiluminescence detector and a computer. Four PTFE connecting tubes with an inner diameter of 0.8mm connect the multi-channel peristaltic pump (P1, P2), micro-injection pump (P3), eight-way injection valve and sensor. , rapid immune response, the formed pre-incubation complex is transported by the microsyringe pump (P3) and flows out through the immunosensor, the carrier fluid, washing solution and regeneration solution are transported by the peristaltic pump (P1), and the luminescent substrate is transported by the peristaltic pump (P2) After mixing, it is injected through the eight-way injection valve and passed thro...

Embodiment 3

[0037] Embodiment 3: Chemiluminescence immunoassay method:

[0038] All analysis steps are programmed and automatically controlled by computer. The regeneration solution is 0.1M amino acid / hydrochloric acid buffer, pH2.0; the washing buffer is 0.01M phosphate buffer containing 0.05% Tween-20, pH7.0; the chemiluminescent substrate is 0.5mmol / l luminol , A mixed solution of 4mmol / l hydrogen peroxide and 0.6mmol / l p-iodophenol. The specific analysis process is as follows:

[0039] (1) Add the antigen-containing sample to be tested and the tracer antibody labeled with horseradish peroxidase into the incubation tube, blow nitrogen gas to generate microbubbles, and incubate at room temperature for 10 minutes under the action of microbubbles.

[0040] (2) Use a microsyringe pump (P3) to pass the antigen-antibody complex through the immunosensor at 0.025 mL / min.

[0041] (3) Pass the flushing solution through the immunosensor at a flow rate of 2.0mL / min through the peristaltic pump...

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Abstract

The invention relates to a chemiluminescent immunological detection system with microbubble-accelerated immunoreaction, which comprises a solution conveying system, a micro-bubble incubation device, an immunosensor, a chemiluminescent detector and a computer. The conveying system consists of peristaltic pumps (P1, P2), a micro syringe pump (P3), a connecting pipe and an eight-way syringe valve; and the immunosensor is made of colorless transparent glass pipes filled with antibody-modified carboxyl-activated resin microballoons and is placed above an optical window of the chemiluminescent detector. A sample to be detected and an enzyme labeled antibody are added into an incubation pipe, nitrogen is introduced to the incubation pipe to generate micro bubbles, accelerate the immunoreaction and reduce the preincubation time, and the micro syringe pump is used for injecting an incubation complex into the immunosensor to form a sandwiched immune complex. Washing liquor is introduced in by the peristaltic pump (P1) to rinse uncombined excessive antibodies, small amount of chemiluminescent substrate of horseradish peroxidase is injected into the mixture by the peristaltic pump (P2), then carrying current is passed in by the peristaltic pump (P1) to carry a substrate zonal belt into the immunosensor and generate chemiluminescent signals, and the concentration of the sample is detected by the signal intensity. The reproduced liquid is introduced into the mixture by the peristaltic pump (P1) after the detection and a reproducible immunosensor performs the next analysis flow. The detection system has the advantages of low cost, quickness, high sensitivity, reproducibility and good precision, and is suitable for the fields of environmental monitoring, clinical diagnosis, food security, and the like.

Description

1. Technical field [0001] The present invention is a chemiluminescent immunodetection system developed by combining the accelerated immune response of microbubbles with a novel immunosensor made of antibody-modified resin microbeads, especially a rapid immune analysis method for accelerated immune responses by microbubbles; the sensor also relates to the use of A detection system for one-component chemiluminescent immunoassays. 2. Background technology [0002] As a highly selective and sensitive analytical method, immunoassay has been widely used in clinical diagnosis, environmental monitoring, food safety and other fields. Immunoassays are divided into homogeneous and heterogeneous immunoassays, the latter being widely used because of higher sensitivity. Chemiluminescence analysis is also an analytical technique that has developed rapidly in recent years. Its instruments are cheap, easy to operate, and environmentally friendly. It is currently one of the most sensitive an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/546G01N21/76
Inventor 严枫杨占军鞠熀先
Owner JIANGSU CANCER HOSPITAL
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