Gold-labeled test strip for rapid detection of chromium ions as well as preparation method and application thereof
A technology of gold standard test paper and chromium ions, which is applied in the fields of immunology and hygienic inspection, can solve the problems of inability to operate on site, small amount of sample screening, cumbersome and time-consuming detection, etc., and achieves easy operation and control, low detection cost, and high detection efficiency. The effect of simple method
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Embodiment 1
[0051] Embodiment one, Cr 3+ Preparation and Identification of Ionic Artificial Immunization Antigen
[0052] (1) Antigen preparation
[0053] (1) Weigh 10 mg chelating agent ITCBE and dissolve it in 1 mL DMSO to form a metal chelating agent solution; weigh 18.1 mgCrCL 3 ·6H 2 O was dissolved in 1 mL of HEPES buffer with a concentration of 10 mmol / L and a pH value of 8.0 to form Cr 3+ solution; the metal chelating agent solution and Cr 3+ The solution was mixed and the pH value was adjusted to 7.0 with NaOH, and then placed on a shaker at room temperature for 12 h to form Cr 3+ - ITCBE chelate hapten solution;
[0054] (2) Weigh 20 mg of carrier protein BSA and dissolve in 1 mL of HEPES buffer with a concentration of 10 mmol / L and a pH value of 8.0 to form a carrier protein solution;
[0055] (3) Take 1 mLCr 3+ -ITCBE chelate hapten solution was added to 1 mL carrier protein solution, and the pH value was adjusted to 9.0 with NaOH, reacted on a shaker at room temperatur...
Embodiment 2
[0060] Embodiment two, anti-Cr 3+ Preparation and identification of ionic monoclonal antibodies
[0061] (1) Anti-Cr 3+ Ionic monoclonal antibody preparation
[0062] (1) Immunization of animals. with Cr 3+ -ITCBE-BSA was used to immunize 5 Balb / c mice at a dose of 50 μg·0.2 mL / mouse, and injected subcutaneously at multiple points on the back. For the first immunization, Cr was diluted with sterile phosphate buffered saline (PBS) 3+ -ITCBE-BSA, emulsified with an equal volume of complete Freund's adjuvant (CFA); for booster immunization, dilute Cr with sterilized PBS 3+ -ITCBE-BSA, emulsified with an equal amount of Freund's incomplete adjuvant (IFA), the second immunization was performed 3 weeks after the first immunization, and then immunized once every 3 weeks, a total of 5 immunizations, 10 days after the third immunization Blood was collected by docking the tail, placed in a water bath at 37°C for 30 min, placed overnight at 4°C, centrifuged at 800 r / min at 4°C for ...
Embodiment 3
[0073] Embodiment three, the preparation of chromium ion immunogold standard test paper
[0074] (1) Colloidal gold solution preparation. Prepared by the citrate reduction method, take a 1000 mL Erlenmeyer flask, add 975 mL of double distilled water to boil, add 15 mL of trisodium citrate and continue heating for 5 min, add 10 mL of chloroauric acid with a mass concentration of 1%, and continue heating to the solution It is orange-red, and after cooling to room temperature, store it at 4°C for later use;
[0075] (2) Preparation of gold-labeled antibody. Centrifuge the monoclonal antibody solution to be labeled at 10,000 r / min for 30 min, and use 0.1 mol / L K 2 CO 3 Adjust the pH value of the colloidal gold solution to 8.2, add the monoclonal antibody solution with a concentration of 0.2 mg / mL dropwise under magnetic stirring, incubate at room temperature for 20 min, centrifuge at 10,000 r / min for 30 min, discard the supernatant, and use the precipitate for 0.02 mol / L Na 2...
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