Kit and method for detecting hepatitis B virus surface antigen (HBsAg)
A technology for hepatitis B virus and detection reagents, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of detecting HBsAg without metalloporphyrin complexes, and achieves overcoming high non-specific background signal and low labeling rate, The effect of reducing systematic errors and improving detection sensitivity
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Embodiment 1
[0027] Embodiment 1 Novel water-soluble A with high labeling rate 3 Preparation of B-type metalloporphyrin-labeled antibody
Embodiment 2A3
[0028] Add 10 mg of N-hydroxysuccinimide (NHS) into 1 mL of aqueous solution containing 5 mg of metalloporphyrin, stir magnetically at 20°C for 10 min, then add 4 mL of aqueous solution containing 2.0 mg of hepatitis B virus surface antibody (HBsAb), and continue at 20°C Stir for 3 hours, the product was centrifuged at 3000r / min at 4°C for 20 minutes, the supernatant was salted out with 10 g of solid ammonium sulfate, and the precipitate after salting was dialyzed with phosphate buffer (pH 7.0) for 1 hour until the permeate was free. Color and freeze-dry to separate metalloporphyrin-labeled HBsAb with a high labeling rate, which is the detection reagent for HBsAg. Example 2A 3 Chemiluminescence immunodetection of HBsAg labeled with type B metalloporphyrin HBsAb
[0029] (1) Coating: Use pH 7.0, 0.1mol / L phosphate buffer solution to dilute 125 μL 1.0 mg / mL HBsAb aqueous solution to 50.0 mL, prepare 2.5 μg / mL coating solution, add 100 μL per well to 96-well white Place in a cl...
Embodiment 3
[0033] Example 3A 3 Chemiluminescence immunodetection of HBsAg labeled with type B metalloporphyrin HBsAb
[0034] (1) Coating: Dilute 50 μL of 1.0 mg / mL HBsAb aqueous solution to a volume of 50.0 mL with pH 7.4, 0.1 mol / L phosphate buffer solution to prepare 1.0 μg / mL coating solution, and add 200 μL to each well of 96-well white In a bottom-transparent polystyrene microwell plate, place at 37°C for 60 minutes, discard the coating solution, add 200 μL of the above-mentioned phosphate buffer solution containing 5% bovine serum albumin (BSA) to each well as a blocking solution, block at 37°C for 1 hour, Wash with pH 7.4, 0.1mol / L phosphate buffer solution, soak for 3 minutes each time, wash the plate 3 times, vacuum dry at 30°C and put it in a sealed bag for vacuum storage to prepare a microwell plate coated with HBsAb;
[0035] (2) Adding samples: divide the microwell plate coated with HBsAb prepared in step (1) into a standard group and a sample group to be tested, and the c...
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