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Novel ELISA biochemical optical disk detection system

An enzyme-linked immunosorbent assay and enzyme-linked immunosorbent technology, applied in the direction of analytical materials, instruments, etc., can solve problems such as slow development, and achieve the effects of powerful functions, controllable flow rate, and improved detection sensitivity.

Active Publication Date: 2014-09-03
ZHEJIANG PUSHKANG BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The development of this field in China is relatively slow. At present, there is no enterprise engaged in the research and development of automatic biochemical detection discs for ELISA protein detection.

Method used

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  • Novel ELISA biochemical optical disk detection system
  • Novel ELISA biochemical optical disk detection system
  • Novel ELISA biochemical optical disk detection system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Design of ELISA CD-ROM

[0026] figure 1 It is the schematic diagram of the design of the enzyme-linked immunosorbent biochemical detection CD-ROM. Antibodies and antigens are placed in reagent tanks 1 and 1', markers and substrates are placed in reagent tanks 3 and 5, respectively, and reagent tanks 2 and 4 are used for washing solutions. The detection disc designed by the present invention contains many microfluidic design components, for example, the design of valves, injection holes, reagent tanks, detection areas, and waste liquid collection pools. One ELISA disc can detect 12, 24 or 48 samples, or one CD can perform 12, 24 or 48 ELISA tests on one sample.

Embodiment 2

[0027] Embodiment 2: Improvement and flow rate control of super-hydrophobic valve

[0028] Using amorphous fluoropolymer (CYTOP 805A) and polyaniline nanofibers to cover the surface of polymethyl methacrylate (PMMA), the contact angle and SEM photos before and after protein adsorption were compared, see figure 2 . The results showed that the PMMA surface modified by CYTOP 805A-polyaniline nanofibers was superhydrophobic, and the protein adsorption had little effect on the valve contact angle (only decreased from 175° to 167°). The snapshot photos after loading the dye solution show that the CYTOP-polyaniline nanofiber modified valve can effectively control the programmed burst release and flow rate of the liquid. Compared with other commercial surface modification technologies, the surface modification of CYTOP-polyaniline nanofibers is simple to operate, requires almost no equipment, low cost, uniform distribution of modified surface, smaller intra-batch and inter-batch dif...

Embodiment 3

[0030] Example 3: Optimization of primary antibody adsorption

[0031] as indicated Figure 4 As shown, polyethyleneimine PEI was used as a bridging agent, tyrosinase (TR) catalyzed the reaction, and protein A was immobilized on the PMMA surface for the detection of γ-interferon antibody. The results show that due to the spatial distance formed by the PEI chains, the antibodies have enough space to arrange in order and maintain good activity. Compared with the traditional glutaraldehyde cross-linking method, not only the binding rate and detection sensitivity of the antibody are improved (by 5-10 times), but also the detection signal intensity is increased by 62 times ( Figure 5 ).

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Abstract

The invention discloses a novel ELISA biochemical optical disk detection system. The detection system comprises an ELISA optical disk and a detecting device. The ELISA optical disk combines with micro processing technology, centrifugal fluid technology and enzyme-linked immunosorbent assay; a microfluidic channel, a liquid storage chamber and a capillary valve system are recorded in a special plastic optical disk; and when centrifugal force generated by rotation of optical disk is greater than capillary force at the valve, liquid in the liquid storage chamber releases according to ELISA operation procedures, so as to complete enzyme linked immunosorbent assay reaction. The system has characteristics of automatic detection, small reagent consumption, low cost, fastness, efficiency, high signal sensitivity, simple operation, and convenient data processing and transmission. The system can be applied to prediction, diagnosis, treatment and monitoring of various clinical diseases, and the fields of food safety assessment and environmental monitoring.

Description

technical field [0001] The invention relates to a novel ELISA biochemical disc detection system. The system combines the advantages of microprocessing technology, centrifugal fluid technology and ELISA technology, and can be widely used in the fields of biomedicine, clinical diagnosis, food safety and environmental detection. Background technique [0002] The commonly used enzyme-linked immunoassay assay (ELISA) operates on a traditional 96-well plate and consists of a series of mixing, incubation and washing processes. The first step is to add the antibody (primary antibody) to the well plate, incubate for a certain period of time to allow the antibody to adsorb to the solid surface, and then wash to remove the unadsorbed antibody; the second step is to add blocking protein solution to occupy the solid that is not adsorbed by the primary antibody. surface to reduce non-specific adsorption of proteins or antibodies. The third step is to add samples or standards containing k...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N35/00
Inventor 余波李利何宏燕钱江张卫
Owner ZHEJIANG PUSHKANG BIOTECHNOLOGY CO LTD
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