Detection micro particle of thyroid stimulating hormone and preparation and application thereof
A technology of thyroid-stimulating hormone and microparticles, which is applied in biological testing, measuring devices, material inspection products, etc., and can solve problems such as expensive instruments and complex automation design
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Embodiment 1
[0097] Preparation of luminescent particles coated with TSHAb in embodiment 1
[0098] Preparation method of antibody and luminescent particles:
[0099] 1) Suspension treatment of luminescent particles: absorb a certain amount of carboxyl luminescent particles and centrifuge in a high-speed refrigerated centrifuge, discard the supernatant, add a certain amount of MES buffer, ultrasonically break until the particles are resuspended, add MES buffer to adjust the luminescent particles Concentration to 100mg / ml.
[0100] 2) Antibody treatment: TSH antibody was dialyzed against 0.05M MES buffer at pH 6.0 (hereinafter referred to as MES buffer). After the dialysis, the concentration was measured and adjusted to 8 mg / ml.
[0101] 3) MES buffer, 100mg / ml luminescent microparticle suspension (MES buffer) and 8mg / ml anti-TSH (MES buffer) and mixed at a volume ratio of 1:1:1, and mixed quickly , to obtain the reaction solution.
[0102] 4) Prepare 40mg / ml EDAC solution with MES buffe...
Embodiment 2
[0150] Example 2 Preparation of biotin-labeled antibody
[0151] Preparation:
[0152] 1) Antibody treatment: dialyze anti-TSH antibody in 0.1M NaHCO 3 solution, the antibody concentration was determined and adjusted to 1 mg / ml.
[0153] 2) Prepare 16.17mg / ml Biotin solution with DMSO.
[0154] 3) Labeling: Take the processed 1 mg / ml anti-AFP antibody labeled antibody and the prepared Biotin solution, mix them according to the volume ratio of 72:10000, and mix them uniformly quickly. Stand at 4°C for 12-16 hours.
[0155] 4) Dialysis: Dialyze the reacted biotin-labeled antibody against biotin-labeled dialysis buffer (pH 8.0).
[0156] 5) Aspirate the dialyzed biotinylated antibody and transfer it to a clean centrifuge tube, and take a sample to determine the antibody concentration. Adjust the concentration of qualified biotin-labeled antibody to 0.5mg / ml.
[0157] React antibodies with Biotin in different ratios and detect:
[0158] The detection method of each paramete...
Embodiment 3
[0191] Example 3 Preparation of Photosensitive Microparticles Coated with Avidin
[0192] Photosensitive particles: use photosensitive particles with a particle size of 220±40nm (PentaTek, USA)
[0193] Preparation:
[0194] a. Treatment of photosensitive particle suspension: absorb a certain amount of photosensitive particles and centrifuge in a high-speed refrigerated centrifuge, discard the supernatant, add a certain amount of MES buffer, ultrasonically sonicate on the ultrasonic cell disruptor until the particles are resuspended, and then add MES buffer Adjust the photosensitive particle concentration to 100mg / ml.
[0195] b. Preparation of avidin solution: weigh a certain amount of Avidin, add MES buffer to dissolve to 8mg / ml.
[0196] c. Mixing: Mix the treated photosensitive microparticle suspension, 8 mg / ml Avidin and MES buffer at a volume ratio of 2:5:1, and mix quickly to obtain a reaction solution.
[0197] d. Reaction: Prepare 25mg / ml NaBH in MES buffer 3 CN s...
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