A functionalized biological multilayer pore membrane immune carrier, preparation method and application
A functionalized and porous technology, applied in the field of immunodetection, can solve the problems of not being widely used, limited antibody storage conditions, limited adsorption capacity, etc., and achieve the effect of firm specific binding, firm adsorption capacity, and enhanced reaction sensitivity.
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Embodiment 1
[0085] like image 3 As shown, the preparation of functionalized biological multilayer pore membrane immune carrier includes the following steps:
[0086] 1) Preparation of biological multilayer porous membrane
[0087] First, the silkworm cocoon (silk fabric) is cut from the long axis direction to remove silkworm chrysalis, miscellaneous silk and other impurities. Wash it twice with pure water, soak it in 100°C water for 15 minutes, take it out and dry it at room temperature. Then, a 5-9mm circular biomultilayer pore membrane was punched with a hole puncher. The silkworm slices were soaked in the treatment solution and incubated in a water bath at 50°C for 2 hours. Use a vacuum dryer to remove air bubbles once, all circular biological multilayer pore membranes sink to the bottom of the treatment solution, continue to incubate in a 50°C water bath for 2 hours, and wash twice with the lotion to prepare the desired biological multilayer pore membranes.
[0088] 2) Preparatio...
Embodiment 2
[0091] like Figure 4 As shown in the figure, the preparation of the biological multi-layer pore membrane immune carrier for the detection of human ABO and Rh(D) blood group positive typing includes the following steps:
[0092] 1) Prepare a functionalized biological multilayer pore membrane immune carrier according to the method described in Example 1;
[0093] 2) Functionalized biological multilayer pore membrane immune carrier conjugated with red blood cell blood group antibody
[0094] Commercially available monoclonal anti-A and anti-B with a titer of 128 were added to the functionalized biological multilayer pore membrane immune carrier obtained in step 1), overnight at 4°C. Air-dry at room temperature at 20-25° C., and store at 4° C. for later use to prepare a biological multilayer pore membrane immune carrier for human ABO blood group detection.
[0095] The final concentration of monoclonal anti-human IgG 20μg / ml was added to another part of the biological multi-lay...
Embodiment 3
[0097] The preparation of the biological multilayer pore membrane immune carrier for the detection of human ABO blood group reverse typing includes the following steps:
[0098] 1) Prepare the functionalized biological multilayer pore membrane immune carrier according to the method described in Example 1.
[0099] 2) Preparation of sensitized erythrocyte blood group antigen
[0100] Take fresh A-type, B-type and O-type red blood cells, respectively prepare a 50% red blood cell suspension with normal saline, add equal volumes of monoclonal anti-A, anti-B and anti-H (titer 8) respectively, and react at room temperature for 5 minutes At this time, red blood cells will appear sand-like agglutination (agglutination strength 1+), add 5 times the volume of hypotonic solution (0.1%-0.7% NaCl solution), stand at room temperature for 1 minute, and then centrifuge at 12000rpm for 5 minutes. The supernatant was discarded, and the sensitized erythrocyte blood group antigen was collected a...
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