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Protein chip fluorescence detection method

A protein chip and protein fluorescence technology, applied in the field of biomedicine, can solve the problems of large detection area, complex processing technology, uneven capture, etc., and achieve the effect of uniform capture, simple operation and low cost

Inactive Publication Date: 2019-05-17
江苏医联生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional protein chip solid-phase carrier has a large detection area, complex processing technology, and uneven capture

Method used

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  • Protein chip fluorescence detection method
  • Protein chip fluorescence detection method
  • Protein chip fluorescence detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] A protein chip fluorescence detection method, comprising:

[0039] Step 1: Perform PL treatment on the surface of the glass sheet; specifically include the following steps:

[0040] Step (1) respectively use acetone, isopropanol and deionized water to ultrasonically clean the glass sheet for 5 minutes;

[0041] Step (2) After drying the glass sheet with nitrogen, as figure 1 As shown, the glass piece was immersed in NaOH solution with a concentration of 2.5M in water and reacted at room temperature for 2h;

[0042] Step (3) Rinse the glass sheet with running water and dry it with nitrogen gas, then react with the surface of the glass sheet with 0.1% PL solution at room temperature for 2 hours, and obtain a glass carrier with PL attached to the surface after cleaning;

[0043] Step 2: Carry out ELISA experiment on the glass plate that this surface has been processed by PL; This process comprises the following steps:

[0044] Step (1) adding 20 μg / ml cardiac troponin I...

Embodiment 2

[0052] A protein chip fluorescence detection method, comprising:

[0053] Step 1: Use a PDMS mold with a hole in the center to treat a specific area on the surface of the glass sheet with PL; PL is the abbreviation of poly-L-lysine, poly-l-lysine; specifically includes the following steps:

[0054] Step (1) Use a glass rod to mix the PDMS solution (PDMS is polydimethylsiloxane) with a ratio of 10:1 evenly, and then vacuumize for 1 hour to remove the air bubbles;

[0055] Step (2) Slowly pour the PDMS mixture onto a clean Si sheet, and heat at 80°C for 1 hour to solidify;

[0056] Step (3) Use a knife to cut the PDMS cured block into small cured blocks of 2cm*2cm, and take 3 of the small cured blocks and punch 1mm, 2mm and 3mm diameter circular penetrating small blocks in the center of them with a puncher. The injection port of the solidified block, to obtain three PDMS molds; as figure 2 shown;

[0057] Step (4) Use acetone, isopropanol and deionized water to ultrasonicall...

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Abstract

The invention relates to a protein chip fluorescence detection method. The method comprises the following steps: step one, performing PL treatment on a surface of a glass sheet; step two, performing ELISA test on the glass sheet after performing the PL treatment; the step one comprises the step (1), ultrasonically washing the glass sheet by using acetone, isopropanol and deionized water in order;step (2) blow-drying the glass sheet by using nitrogen, immersing the glass sheet in the NaOH solution with water concentration of 2.5M, reacting for 2h at room temperature; step (3) cleanly flushingthe glass sheet by using running water, and blow-drying by using the nitrogen, and enabling 0.1% of PL solution to react with the surface of the glass sheet for 2h at the room temperature, and obtaining a glass carrier attached with the PL on the surface after completely cleaning the glass sheet. The method provided by the invention is simple in operation and low in cost, the detection size is adjustable, and the fluorescence detection performance of the low-concentration protein is provided; an operator can obtain the detection region with required size by firstly preparing a required PDMS mold and then chemically treating the glass carrier surface at a specific region, and the capturing is uniform.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a protein chip fluorescence detection method. Background technique [0002] Protein chip technology has potential broad application prospects in the field of biomedicine. The protein chip can simultaneously detect the content of all proteins in biological samples that may be related to a certain disease or environmental factor damage, that is, the phenotypic fingerprint. Phenotype fingerprints are also of great significance for monitoring the process or prediction of diseases and judging the effect of treatments. Traditional protein chips generally carry out special chemical treatment on the whole solid phase carrier, and then immobilize known protein molecules on it to capture the protein to be tested that can specifically bind to it, so as to realize the detection and analysis of biomolecules. Provide strong technical support in areas such as drug screening. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/531G01N33/533G01N33/543G01N33/577G01N33/68
Inventor 杨文婷曹臻陈亮
Owner 江苏医联生物科技有限公司