Chromium ion rapid detection gold standard test paper and its preparation method and application
A technology of gold standard test paper and chromium ions, which is applied in the fields of immunology and sanitary inspection, can solve the problems of cumbersome and time-consuming detection, small sample screening volume, and inability to operate on-site, and achieve simple detection methods, low detection costs, and easy operation and the effect of controlling
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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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