Microsphere lyophilized preparation coupled with proteins and preparation method and preservation mode thereof
The technology of a freeze-dried preparation and a freeze-drying protective agent is applied in the field of protein-coupled microsphere freeze-dried preparation and its preparation, which can solve the problems of short shelf life and poor stability, achieve simple formula, improve stability and reduce transportation. effect on storage costs
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Embodiment 1
[0056] The preparation of embodiment 1 lyoprotectant
[0057] Preparation method: weigh 0.2g sodium dihydrogen phosphate, 2.9g disodium hydrogen phosphate dodecahydrate, 0.2g potassium chloride, 8g sodium chloride, 30g trehalose, 40g mannitol, 10g polyvinylpyrrolidone , 5g of dextran, add 800mL of purified water and stir to dissolve, measure 50mL of donkey serum, 300μL of Proclin300, add to the solution and mix well, use purified water to make the volume of the prepared solution to 1L, finally, use 0.22μm filter membrane to filter the solution.
[0058] Wherein, the weight ratio of trehalose: polyvinylpyrrolidone: dextran is 6:2:1. The components and contents of the prepared lyoprotectant are shown in Table 1.
Embodiment 2
[0068] The freeze-dried preservation of the microsphere of embodiment 2 coupling SOX2 antigenic protein
[0069] (1) Preparation of microspheres coupled with SOX2 antigen protein
[0070] The SOX2 antigen protein (GenScript Company) with c-Myc tag at the N-terminus was coupled to fluorescently encoded MagPlex magnetic microspheres from Luminex Company. The coupling method was operated in accordance with the instructions of the xMAP Coupling Kit (Luminex Antibody Coupling Kit, Cat. No. 40-50016), as follows:
[0071] 1) Activation of microspheres
[0072] a) take out For microspheres, vortex and sonicate for about 20s to fully mix and resuspend the microspheres. According to the number of microspheres required for coupling, draw an appropriate volume of microsphere suspension (about 5×10 6 microspheres) in a centrifuge tube.
[0073] b) Place the centrifuge tube on the magnetic separator for 30-60 seconds to separate the microspheres from the liquid. Carefully remove the...
Embodiment 3
[0091] The freeze-dried preservation of the microsphere of the monoclonal antibody of embodiment 3 coupling carcinoembryonic antigen (CEA)
[0092] In this example, the monoclonal antibody of carcinoembryonic antigen (CEA) was coupled to the microspheres, and the CEA monoclonal goat antibody was coupled with fluorescently encoded magnetic microspheres MagPlex commercially available from Luminex in a covalent manner. , using the lyoprotectant prepared in Example 1. Other conditions and steps are with embodiment 2.
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