Colloidal gold immunochromatographic card for detecting bisphenol A (BPA) and preparation method thereof
The technology of colloidal gold and detection card is applied in the field of immunocolloidal gold detection card of bisphenol A and its preparation, and achieves the effects of simple operation, easy popularization and use, and fast detection speed.
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Embodiment 1
[0025] figure 1 It is the structural diagram of the immune colloidal gold detection card of bisphenol A of the present invention, figure 2 It is the cross-sectional structure diagram of the test strip in the immune colloidal gold detection card of bisphenol A of the present invention: among the figure 9 is a PVC glue board; 7 is a sample pad; 8 is a colloidal gold binding pad, and the colloidal gold binding pad is coated with Bisphenol A monoclonal antibody colloidal gold marker; 10 is the coated membrane, that is, nitrocellulose membrane, which is coated with bisphenol A-BSA and rabbit anti-mouse IgG; 11 is absorbent cotton, which is made of absorbent cotton Made of materials such as filter paper.
[0026] A sample pad 7 and a colloidal gold bonding pad 8 are adhered to one end (sample end) of the PVC rubber plate 9 , and the sample pad 7 is lapped on the colloidal gold bonding pad 8 .
[0027] Adhesive coating film 10 in the middle of PVC rubber plate 9. The rabbit anti-...
Embodiment 2
[0030] The preparation of bisphenol A colloidal gold detection card is realized by the following steps:
[0031] 1. Monoclonal antibody: After the hybridoma cell line was conventionally cultured and passaged in vitro, bisphenol A-BSA was injected into the peritoneal cavity of BALB / C mice, the ascites was collected, and the monoclonal antibody was purified by affinity chromatography;
[0032] 2. Preparation of colloidal gold solution: Take a 1 L Erlenmeyer flask, add 495 mL of ultrapure water, and then add 1% chloroauric acid (HAuCl 4 ·3H 2 O) 5 mL, prepared into 500 mL 0.01% chloroauric acid aqueous solution, heated to boil, then added 1% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) Solution 5-7 mL, continue to stir and heat, when the color of the solution turns completely transparent purple red, stop heating after maintaining for 5 minutes, add water to the original volume, cool to room temperature, and store at 2-8°C for later use;
[0033] 3. Antibody pretreatm...
Embodiment 3
[0041] Minimum detection limit test
[0042] Bisphenol A minimum detection limit test
[0043] Accurately weigh 0.500 g of bisphenol A standard substance, dissolve it in ultrapure water and dilute to 100 mL, mark it as A 1 solution, take A 1 Add 1 mL of the solution to a volumetric flask and add ultrapure water to 1000 mL to form A 2 solution, take A 2 Add 1 mL of the solution to a volumetric flask and add ultrapure water to 1000 mL to form A 3 solution, then A 3 The concentration of the solution is 5 ng / mL (5 ppb). take A 3 Put 1 mL of the solution in 4 test tubes, and label them 1~4 respectively, add 9 mL, 4 mL, 1 mL, and 0 mL of ultrapure water to the test tubes respectively, and prepare the bisphenol A standard solution with concentrations of 0.5 ng / mL, 1 ng / mL, 2.5 ng / mL, 5 ng / mL; take another empty test tube, add 5 mL ultrapure water, and mark it as No. 5 test tube. Take the liquid in the above test tubes 1-5 as samples and add them dropwise to the bisphenol A t...
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