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Dry method photic stimulation chemiluminescence immunoassay reagent kit and preparation and application thereof

A photo-excited chemiluminescence and immunodetection technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc. Effect

Active Publication Date: 2010-06-23
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the existing publications, regarding the application of the above technologies, the particles and related reagents are stored and transported under liquid conditions. During the detection operation, it is necessary to add the relevant reagents to the reaction vessel one by one manually or by using an automatic sampling platform. The process is cumbersome, there are multiple steps of adding samples, and professionals are required to perform related operations

Method used

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  • Dry method photic stimulation chemiluminescence immunoassay reagent kit and preparation and application thereof
  • Dry method photic stimulation chemiluminescence immunoassay reagent kit and preparation and application thereof
  • Dry method photic stimulation chemiluminescence immunoassay reagent kit and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Signal detection of quality control microparticles

[0042] (1) Preparation of quality control particles:

[0043] a. Preparation of biotinylated γ-globulin (BGG)

[0044] with 0.1M NaHCO 3 BGG (purchased from Pel-Freez Biological) was formulated into a 1mg / ml solution, and Biotin-X-X-NHS (N-hydroxysuccinimide modified biotin) was prepared using DMSO (dimethylformamide), manufacturer: SIGMA, product No.: B3295) solution to 16.172mg / ml, take 5.4ul Biotin-X-X-NHS to 1mg BGG solution, mix well and place overnight at 4°C. The purified protein was dialyzed against 100 mM phosphate buffer (pH 7.0).

[0045] b. Use the above-mentioned labeled protein to coat aldehyde-modified luminescent particles

[0046] Add 12.5 μl of 1% Tween-20, 0.05 mg of the dialyzed purified protein obtained in step 1 above, and 10 μl of NaCNBH to 1 mg of aldehyde-modified luminescent particles (PentaTek, USA). 3 (25mg / ml), add 0.1M pH6.0 MES to a total volume of 200μl, and incubate at ...

Embodiment 2

[0063] Embodiment 2: the detection of HBsAg (hepatitis B surface antigen)

[0064] (1) Preparation and preparation of biotin-labeled surface antibody solution

[0065] Preparation: a. Dialyze HBsAb in 0.1M NaHCO 3 solution, the antibody concentration was measured and adjusted to 1 mg / ml. b. Prepare 16.17mg / ml Biotin solution with DMSO. c. Labeling: Take the processed 1 mg / ml HBsAb labeled antibody and the prepared Biotin solution, mix them according to the volume ratio of 72:10000, and mix them evenly quickly. Stand at 4°C for 12-16 hours. d. Dialysis: Dialyze the reacted biotin-labeled antibody against biotin-labeled dialysis buffer (pH 8.00). e. Aspirate the dialyzed biotinylated antibody and transfer it to a clean centrifuge tube, and take a sample to determine the antibody concentration. Adjust the concentration of qualified biotin-labeled antibody to 0.5mg / ml.

[0066] Preparation: Dilute the biotin-labeled surface antibody with biotin buffer (0.02M MES, 1% BSA, 1% ...

Embodiment 3

[0078] Embodiment 3: the preparation of cTnI (cardiac troponin I) dry type diagnostic reagent

[0079] (1) Preparation and preparation of biotin-labeled anti-cardiac troponin I antibody solution

[0080] Preparation: a. Dialyze anti-myocardin I antibody in 0.1M NaHCO 3 solution, the antibody concentration was measured and adjusted to 1 mg / ml. b. Prepare 16.17mg / ml Biotin solution with DMSO. c. Labeling: Take the processed 1 mg / ml HBsAb labeled antibody and the prepared Biotin solution, mix them according to the volume ratio of 72:10000, and mix them evenly quickly. Stand at 4°C for 12-16 hours. d. Dialysis: Dialyze the reacted biotin-labeled antibody against biotin-labeled dialysis buffer (pH 8.00). e. Aspirate the dialyzed biotinylated antibody and transfer it to a clean centrifuge tube, and take a sample to determine the antibody concentration. Adjust the concentration of qualified biotin-labeled antibody to 0.5mg / ml.

[0081] Preparation: Dilute the biotin-labeled ant...

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Abstract

The invention relates to an immunoassay reagent kit, and discloses a dry method photic stimulation chemiluminescence immunoassay reagent kit and preparation and application thereof. The dry method photic stimulation chemiluminescence immunoassay reagent kit of the invention comprises a reaction vessel of an antigen or an antibody fixed with dry detection particles and dry biotin marks, wherein, the detection particles are luminous particles of the envelope antigen or the antibody. The kit of the invention can be used for quantitative or qualitative detection of the antigen or the antibody to be detected.

Description

technical field [0001] The invention relates to an immunoassay kit, in particular to a dry-method light-excited chemiluminescence immunoassay kit, its preparation and its use. Background technique [0002] Light-induced chemiluminescence technology is a chemiluminescence technology based on polymer particles. It can combine photosensitive particles and luminescent particles within a certain distance to generate ion oxygen energy transfer and send out optical signals, so as to detect the samples to be tested. . Photosensitive particles are polymer particles filled with photosensitive compounds, and luminescent particles are polymer particles filled with luminescent compounds and lanthanide elements. The core principle of photochemiluminescence technology is the short-distance transfer of energy, and the transfer medium is high-energy ionic oxygen. When the red laser irradiates the photosensitive particles, the singlet oxygen ions (4μs) are released, and the propagation dist...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N21/76
Inventor 石晓强王海蛟赵卫国
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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