Fluorescence immunochromatography test strip prepared by taking aggregation-induced emission fluorescent microsphere as beacon carrier
A technology of aggregation-induced luminescence and fluorescence immunochromatography, which is applied in the fields of medical testing and food safety testing, can solve the problems of preparing fluorescent microspheres without tetramethyl tetrastyrylate material, and achieves good fluorescence characteristics and detection sensitivity. High, high-capacity effects
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Embodiment 1
[0036] Example 1: Preparation of aggregation-induced luminescence fluorescent microspheres (preparation of TCBPE dyes)
[0037] 1. Prepare the dye preparation solution: Weigh TCBPE (the synthesis method of TCBPE comes from the document Rigidifying Fluorescent Linkers by Metal-Organic Framework Formation for Fluorescence Blue Shift and Quantum Yield Enhancement Journal of the American Chemical Society 2014 136 (23), reported in 8269-8276), PMMA and PMAO is dissolved in chloroform, so that the final concentrations of TCBPE, PMMA, and PMAO dissolved in chloroform are 30, 50, and 25 mg / mL respectively;
[0038] 2. Preparation of microemulsion: Take 2400 μL of the above prefabricated solution, disperse it in 12 mL of 0.25% SDS solution, and perform ultrasonic emulsification with an ultrasonic breaker for 3 minutes under ice bath conditions, and the emulsification power is 600W;
[0039] 3. Solidify into balls: put the above emulsion in a rotary evaporator, and evaporate until there...
Embodiment 2
[0040] Example 2: Preparation of a competitive immunochromatographic test strip for the detection of 25-hydroxyvitamin D (25-OH-VD) prepared using aggregation-induced luminescence fluorescent microspheres as a beacon carrier
[0041] 1. Preparation process of immunochromatographic test strips
[0042] 1. Preparation of nitrocellulose membrane;
[0043] (1) Preparation of 25-OH-VD artificial antigen (25-OH-VD-OVA):
[0044] The coupling method is mixed acid anhydride method, the coupling protein is ovalbumin (OVA), and the coupling ratio is 1:80. After coupling, dialysis purification is used to obtain 25-OH-VD-OVA.
[0045] (2) Preparation of detection line and quality control line:
[0046] 25-OH-VD-OVA conjugates and anti-mouse antibodies were coated onto nitrocellulose membranes: dilute the concentration of 25-OH-VD-OVA conjugates with 0.01M PBS (phosphate buffer) at pH 7.5 The concentration of the diluted anti-mouse antibody is 1mg / mL, and the resulting solution is spray...
Embodiment 3
[0061] Example 3: Preparation of a competitive immunochromatographic test strip for the detection of zearalenone prepared using aggregation-induced luminescent fluorescent microspheres as a beacon carrier
[0062] 1. Preparation process of immunochromatographic test strips
[0063] 1. Preparation of nitrocellulose membrane;
[0064] (1) Preparation of ZEN artificial antigen (ZEN-BSA):
[0065] The coupling method is the mixed acid anhydride method, the coupling protein is bovine serum albumin (BSA), and the coupling ratio is 1:100. After coupling, it is purified by dialysis to obtain ZEN-BSA.
[0066] (2) Preparation of detection line and quality control line:
[0067] ZEN-BSA conjugates and anti-mouse antibodies were coated onto nitrocellulose membranes: Dilute the ZEN-BSA conjugates to a concentration of 6 mg / mL with 0.01M pH 7.0 PBS (phosphate buffered saline), and the resulting solution was in Spray on the membrane as a test line; the concentration of the diluted anti-m...
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