Colloidal gold quick detection test paper of retinal binding protein
A technology for detecting test strips and binding proteins, which is applied in the fields of biomedicine and immunoassay, and can solve the problems of long detection time, shelf life of only half a year, and high cost.
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Embodiment 1
[0114] Example 1. Preparation of gold-labeled antibody layer (binding release pad)
[0115] Preparation of colloidal gold
[0116] Using the trisodium citrate method, 0.1% gold chloride was reduced to a colloidal gold solution with 1% trisodium citrate. The quality of the reduced colloidal gold solution is detected by electron microscope observation and 400-700nm wavelength spectrophotometer scanning method, and a colloidal gold solution with a size of about 40nm, uniform size and uniform dispersion is obtained. Adjust the colloidal gold solution to the working concentration (1 mg / ml), and adjust the pH=7.5-8.5.
[0117] Preparation of colloidal gold-labeled anti-urine RBP antibody
[0118] Take 1 ml of colloidal gold solution, adjust the pH to 8.2 with 0.2M potassium carbonate solution, and add 25 μg of commercially available urine RBP antibody (mouse anti-urine RBP monoclonal antibody, purchased from Genetex) or mouse anti-urine RBP prepared by the applicant. Monoclon...
Embodiment 2
[0121] Example 2. Preparation of nitrocellulose membrane with detection line and quality control line
[0122] Get anti-urine RBP polyclonal antibody (prepared according to the method described in 200610117198.7 by this laboratory), adjust its concentration to 1mg / ml (the reagent used to adjust the concentration is PBS), add 2% formaldehyde, and use a film spraying machine (Biodot company, XYZ3050 Model) Spray a test line (t-line, T-line) in the middle of the nitrocellulose membrane, and set the amount of sprayed film to 1 μl / cm.
[0123] Take goat anti-mouse IgG (purchased from SIGMA), adjust the concentration to 2mg / ml (the reagent used to adjust the concentration is PBS), add 2% formaldehyde, and spray it on the middle section of the nitrocellulose membrane with a film sprayer, 0.5cm away from the detection line. For the control line (C-line, C-line), the amount of spray film is set to 1 μl / cm.
[0124] The sprayed film was dried at 37°C for 2 hours, and then blocked with ...
Embodiment 3
[0125] Embodiment 3. Assembly of urine RBP colloidal gold rapid detection test paper
[0126] Assemble the urine RBP colloidal gold rapid detection test strip by the following steps:
[0127] (1) The nitrocellulose membrane with detection line and quality control line obtained in Example 2 is pasted on the PVC base plate;
[0128] (2) Paste the binding release pad prepared in Example 1 on the PVC plate near one end of the test line, so that the binding release pad overlaps with the nitrocellulose membrane at the lower end of the test line by 2 mm;
[0129] (3) Stick water-absorbing glass fiber (sample pad) on the other side of the binding release pad, so that the sample pad overlaps with the binding release pad by 2 mm;
[0130] (4) Paste absorbent paper on the PVC board near one end of the quality control line, so that the absorbent paper overlaps with the nitrocellulose membrane at the top of the quality control line by 2 mm;
[0131] (5) Cut into strips according to the s...
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