Method for preparing prostate cancer antigen immunochromatography test paper based on hydrophobin high-efficiency fixed low-concentration antibody
An immunochromatographic test strip and hydrophobin technology, which is applied in the field of clinical medical detection, can solve the problems of reduced utilization of epitope epitopes, reduced ability to capture antigens, and unsatisfactory detection effects, and can improve the ability to capture antigens and reduce antibodies. The effect of using the content and reducing the cost of testing
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[0032] Preparation of quantum dot immunoprobe: Dissolve water-soluble quantum dots, prostate cancer labeled antibody, and EDC (1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride) in boric acid buffer The solution, suspension culture at room temperature, activate the carboxyl groups of the quantum dots, centrifuge, add the same amount of antibody and repeat the above process, and finally the quantum dots coupled with the antibody are blocked with BSA overnight.
[0033] Assembly of a new type of test strip with the help of hydrophobin to immobilize antibodies: The ordinary test strip is composed of a sample pad, a binding pad, a nitrocellulose membrane, and an absorbent pad. The test strip of the present invention is closely connected from left to right It is a sample pad and a binding pad stacked together, in order to fully bind the antigen and the antibody labeled with quantum dots, such as figure 1 As shown, the number of sample pads in this technical invention is two, wh...
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[0045] Implementation case 1:
[0046] Preparation method of prostate cancer immunochromatographic test strip based on hydrophobin's high-efficiency immobilization of low-concentration antibody; spray 50μg / ml of hydrophobin on the position of the detection line of the nitrocellulose membrane, and the mass fraction ratio of quantum dots and EDC is 1:4000, the mass fraction ratio of quantum dots and antibody is 1:16, BSA blocking solution is 1%, and the concentration of PSA coating antibody is 0.2mg / ml when diluted with 0.01M PBS buffer with PH=7.4 Spray on the hydrophobin layer, their overlapping area is used as the detection line area, drop 45μl of standard antigen with a concentration of 5ng / ml into the sample pool, do three replicates, react for 15 minutes, read out the detection with a quantum dot immunofluorescence analyzer The indications of the line and the quality control line are the T value and the C value, and the average value of the detected signal T / C intensity is 0....
Example Embodiment
[0047] Implementation case 2:
[0048] Preparation method of prostate cancer immunochromatographic test strip based on hydrophobin's high-efficiency immobilization of low-concentration antibody; spray 50μg / ml of hydrophobin on the position of the detection line of the nitrocellulose membrane, and the mass fraction ratio of quantum dots and EDC is 1:4000, the mass fraction ratio of quantum dots and antibody is 1:16, the amount of BSA blocking solution is 1%, and the concentration of PSA coating antibody is 0.2mg / ml when diluted with 0.01M PBS buffer with PH=8 Spray on the hydrophobin layer, their overlapping area is used as the detection line area, drop 45μl of standard antigen with a concentration of 5ng / ml into the sample pool, do three replicates, react for 15 minutes, read out the detection with a quantum dot immunofluorescence analyzer The indications of the line and the control line are the T value and the C value, and the average value of the detected signal T / C intensity i...
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