Colloidal gold immunochromatography test strip for rapidly detecting capsaicinoids as well as preparation method and application thereof
An immunochromatographic test strip and capsaicin technology, which is applied in biological tests, measuring devices, analytical materials, etc., can solve the problems of blurred spiciness level labels, and achieve the effects of simple operation and high detection sensitivity.
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Embodiment 1
[0046] Embodiment 1 Obtaining of anti-capsaicinoid monoclonal antibody
[0047] The anti-capsaicinoid monoclonal antibody is a monoclonal antibody secreted by the hybridoma cell line YQQD8 with the preservation number CCTCCNO.C201534. The specific preparation method:
[0048] The hybridoma cell line YQQD8 was injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, the ascites of the mice was collected, and the antibody was purified by octanoic acid-ammonium sulfate method. Rat ascites, 4°C, centrifuged at 12000r / min for 30min, sucked the supernatant, mixed the obtained ascites supernatant with 3 times the volume of acetate buffer, slowly added n-octanoic acid under stirring, the volume of n-octanoic acid required per milliliter of ascites was 33μL, mix at room temperature for 30min, stand at 4°C for more than 2h; centrifuge at 12000r / min for 30min at 4°C, discard the precipitate, filter the obtained supernatant with double-layer filter paper, add 1 / 10 o...
Embodiment 2
[0075] Embodiment 2: the preparation method of the colloidal gold immunochromatography test paper strip of rapid detection capsaicinoid, the steps are as follows:
[0076] (1) Preparation of absorbent pad
[0077] Cut the absorbent paper to a length of 16mm and a width of 4mm to obtain an absorbent pad;
[0078] (2) Preparation of detection pad
[0079] Coating of the detection line: the capsaicin-like substance-coated antigen, that is, the above-mentioned capsaicin-like substance general-purpose artificial complete antigen-coated antigen, is prepared into a coating solution with a concentration of 0.5 mg / mL, and placed on the upper edge of the nitrocellulose membrane. At the position of 15mm, it was coated horizontally on the nitrocellulose membrane by line spraying method to obtain the detection line. The coating amount of capsaicinoid-coated antigen required for each centimeter of detection line was 0.3 μg, and then at 37°C Dry for 30 minutes;
[0080] The coating buffer...
Embodiment 3
[0100] Embodiment 3: the preparation method of the colloidal gold immunochromatography test strip of rapid detection capsaicinoid substance, the steps are as follows:
[0101] (1) Preparation of absorbent pad
[0102] Cut the absorbent paper to a length of 18mm and a width of 4mm to obtain an absorbent pad;
[0103] (2) Preparation of detection pad
[0104] Coating of the detection line: the capsaicin-like substance-coated antigen, that is, the above-mentioned capsaicin-like substance universal artificial complete antigen-coated antigen, is prepared into a coating solution with a concentration of 0.25 mg / mL, and placed on the edge of the nitrocellulose membrane. At the position of 9mm, it was coated horizontally on the nitrocellulose membrane by line spraying method to obtain the detection line. The coating amount of capsaicinoid-coated antigen required for each centimeter of detection line was 0.15 μg, and then at 37°C Dry for 20 minutes;
[0105] The coating buffer used i...
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