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Method for making sample cell of solid phase white amino reflective layer

A manufacturing method and sample cell technology, which are applied in chemical instruments and methods, material testing products, other chemical processes, etc., to achieve the effects of superior practicability, easy amino acid, high activity and sensitivity

Inactive Publication Date: 2008-04-02
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to improve the sensitivity of ultra-trace immunoassays, develop solid-phase materials suitable for solid-phase time-resolved fluorescence immunoassay applications, and solve technical problems such as replacing transmission with reflection and replacing physical adsorption with chemical covalent bonding, the invention provides a polymer A white opaque nylon layer is fixed on the surface of the styrene microporous plate, which is chemically modified as a solid-phase white amino reflective layer sample pool used in DSLFIA technology.

Method used

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  • Method for making sample cell of solid phase white amino reflective layer
  • Method for making sample cell of solid phase white amino reflective layer

Examples

Experimental program
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Effect test

Embodiment 1

[0051] (1). Add 400 μl of matrix 1 activator to matrix 1 made of PS, leave it at room temperature for 1 minute, discard the solution and immediately add 200 μl of 12% nylon-6 solution dissolved by the middle layer 2 nylon-6 activator, along the central axis Rotate the substrate 1 at a slow speed (40 times / min), so that nylon-6 is quickly, firmly and uniformly combined with the inner surface of the substrate 1 to form a solid-phase white amino reflective layer sample with a nylon-6 film thickness of 0.1-0.2mm The middle layer of the pool is 2 nylon-6 film layers;

[0052] (2). Add 400 μl of 5% triethyloxonium tetrafluoroborate ester dichloromethane solution to the sample pool of the above step (1), leave it at room temperature for 4-10 minutes, discard it, and rinse it twice with dichloromethane , washed with dioxane once;

[0053] 3. Add 200 μl of 0.5 M glutaraldehyde into the sample pool of (2) above, incubate at 37° C. for 15 minutes, and discard. Add 0.2M pH=7.2 phosphate...

Embodiment 2

[0055] (1). Add 400 μl of matrix 1 activator to matrix 1 made of PS, leave it at room temperature for 1 minute, and immediately add 200 μl of 13% nylon-6 solution dissolved with middle layer 2 nylon-6 activator after discarding the solution. Slowly rotate (40 times / min) the substrate 1 along the central axis, so that nylon-6 is quickly, firmly and uniformly bonded to the inner surface of the substrate 1, forming a solid-phase white amino reflective layer of nylon-6 with a thickness of 0.1-0.2mm 2 nylon-6 film layers in the middle layer of the sample pool;

[0056] (2). Add 400l of 4% triethyloxonium tetrafluoroborate ester dichloromethane solution to the sample cell of the above step (1), leave it at room temperature for 4-10 minutes, discard. Rinse twice with dichloromethane , washed with dioxane once;

[0057] 3. Add 200 μl of 0.5 M glutaraldehyde borate buffer solution to the sample cell in the above step (2), incubate at 37° C. for 15 minutes, and discard. Add 0.2M pH=7....

Embodiment 3

[0059] (1). Add 400 μl of matrix 1 activator to matrix 1 composed of PS. Stand at room temperature for 1 minute, discard the solution and immediately add 200 μl of 14% nylon-6 solution dissolved by the middle layer 2 nylon-6 activator. Slowly rotate (40 times / min) the substrate 1 along the central axis, so that nylon-6 is quickly, firmly and uniformly combined with the inner surface of the substrate 1, forming a solid-phase white amino group with a nylon-6 film thickness of 0.1-0.2mm 2 nylon-6 film layers in the middle layer of the reflective layer sample pool;

[0060] (2). Add 400 μl of 3% triethyloxonium tetrafluoroborate ester in dichloromethane to the sample cell in the above step (1), leave it at room temperature for 4-10 minutes, and discard it. Rinse 2 times with dichloromethane, and rinse 1 time with dioxane; The synthetic method of described triethyloxonium tetrafluoroborate is according to the literature: (enzyme is fixed on the chemistry of nylon, biochemistry.J.1...

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Abstract

The invention relates to a making method of solid-phase white amino reflecting layer sample pool, which comprises the following steps: adapting pyridine or acetone to do activating predisposal of the PS surface; dissolving and activating the nylon through acetic acid or m-cresol or pyridine; doing hydrophobic interaction to make the nylon-6 fixation and combination on the inner surface of PS sample pool; disposing the nylon film to make oxygen atom alkylate by TOTFB directly; doing the alkylation acted by glutaral double functional group on the nylon-6 film in the covalent connection with one end of free ammonia on the 'arm' and the other end of free aldehydo in the covalent connection with free ammonia of connection protein as solid-phase amino reflecting layer sample pool. The sample pool can be used repeatedly, which can be used in the solid-phase enzyme method, too.

Description

technical field [0001] The invention belongs to a method for manufacturing a solid-phase white amino reflective layer sample pool. Background technique [0002] Technologies such as solid-phase immunoassay, affinity layer and solid-phase enzymes all need to bind biologically active substances (immunoligands, nucleic acids, enzymes, etc.) to inert macromolecular materials, such as agarose, cellulose, nylon and various plastic surfaces Among them, polystyrene (PS) is the most widely used because of its easy source of raw materials, stable physical and chemical properties, and easy processing and molding. The number and biological activity or affinity of molecules bound to the solid phase surface is one of the factors affecting the detection sensitivity of immunoassays (covalent linkage of haptens, protein and nucleic acid polymers to modified styrene solid phases, Journal of Immunological Methods, Covalent Linking of haptens, protein and nucleic acid to a modified polystyrene...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53B01J20/00
Inventor 潘利华马世盐郭淑英钮倩
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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