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Immunochromatography test paper and preparation method and application thereof

A technology of immunochromatography test paper and streptavidin, which is applied in the field of immunochromatography test paper and its preparation, can solve the problems of limiting the application of immunochromatography technology and sensitivity gap, etc., and achieves the improvement of detection sensitivity, efficiency and sensitivity Effect

Pending Publication Date: 2022-01-25
上海艾瑞德生物科技有限公司
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  • Claims
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AI Technical Summary

Problems solved by technology

However, compared with chemiluminescence detection technology, there is still a large gap in its sensitivity, which limits the application of immunochromatography in the field of diagnosis

Method used

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  • Immunochromatography test paper and preparation method and application thereof
  • Immunochromatography test paper and preparation method and application thereof
  • Immunochromatography test paper and preparation method and application thereof

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preparation example Construction

[0025] The embodiment of the present invention provides the preparation method of the immunochromatography test paper as described in any of the foregoing embodiments, which includes: sequentially overlapping the sample pad, the binding pad, the nitrocellulose membrane and the water-absorbing pad on the bottom plate;

[0026] Step 1: Coating on the binding pad: biotinylated antibodies, and conjugates of antibodies and markers; the biotinylated antibodies and the antibodies in the conjugates are all antibodies that can bind to the analyte .

[0027] In a preferred embodiment, the coating method of the binding pad is as follows: add a biotinylated antibody and a conjugate of the antibody and a marker to the release solution to obtain a binding pad solution; the biotin in the binding pad solution The final concentration of the Fc antibody is 1-5 μg / mL, specifically 1 μg / mL, 2 μg / mL, 3 μg / mL, 4 μg / mL or 5 μg / mL. The final concentration of the conjugate of the antibody and the mar...

Embodiment 1

[0043] The embodiment of the present invention provides an immunochromatographic detection reagent sheet, taking cardiac troponin I (cTnI) as an example, the preparation method is as follows.

[0044] (1) Coating method of nitrocellulose membrane

[0045] Preparation of detection line drawing solution: mix streptavidin and biotinylated antibody (Ab-biotin) in the drawing solution to obtain detection line drawing solution, the final concentration of streptavidin is 2mg / mL, the final concentration of biotinylated antibody was 0.2 mg / mL.

[0046] Prepare independent quality control line antibodies such as goat anti-mouse antibody or goat anti-chicken, DNP-BSA, etc. into 1 mg / mL quality control line drawing solution.

[0047] Fix the stripping solution for the detection line and the stripping solution for the quality control line on the nitrocellulose membrane in a strip shape of 1.0 mm with a gold-spraying film scribing instrument, and make a nitrocellulose package by vacuum dr...

Embodiment 2

[0055] The embodiment of the present invention provides an immunochromatography detection reagent sheet, which is roughly the same as that of embodiment 1, the difference is that the detection line drawing solution is different, and the difference is as follows:

[0056] Streptavidin (SA) was added to the drawing solution to obtain the detection line drawing solution, and the final concentration of streptavidin was 2 mg / mL.

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Abstract

The invention discloses immunochromatography test paper and a preparation method and application thereof, and relates to the technical field of immunochromatography detection.The immunochromatography test paper is characterized in that streptavidin is used for conducting detection line membrane scratching, and biotinylated antibodies are transferred to a combination pad, so that double-antibody sandwich reaction of antigens and antibodies starts from the combination pad, the reaction time is prolonged, and the reaction efficiency is improved; the biotinylated antibody can flow on the nitrocellulose membrane, and the reaction capacity with the antigen is higher than that of the antibody fixed on the detection line, so that the production efficiency of the double-antibody sandwich compound is improved, and the sensitivity is improved. Meanwhile, a capture reaction on the nitrocellulose membrane is converted from an antigen-antibody reaction to a streptavidin-biotin reaction, the reaction efficiency is improved by 3-4 orders of magnitude, and the capture efficiency of a detection line on the membrane is improved, so that the detection sensitivity is improved.

Description

technical field [0001] The invention relates to the technical field of immunochromatography detection, in particular to an immunochromatography test paper and its preparation method and application. Background technique [0002] Immunochromatography is a new type of membrane detection technology based on antigen-antibody specific immune response. In this technology, the strip-shaped fiber chromatography material immobilized with detection line (coated antibody or coated antigen) and quality control line (antibody) is used as the stationary phase, the test solution is used as the mobile phase, and the labeled antibody or antigen is fixed on the binding pad. Capillary action causes the analyte to move on the chromatographic strip, and the analyte and the coated antibody or antigen, labeled antibody or antigen form an immune complex on the detection line. There is a certain relationship between the signal of the marker on the detection line and the analyte. Furthermore, the an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/543
CPCG01N33/558G01N33/543
Inventor 林叶倪晓涛孙博沐舒静
Owner 上海艾瑞德生物科技有限公司
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