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

Active Publication Date: 2020-05-08
GUANGZHOU BIO BLUE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a lyophilized preparation of protein-coupled microspheres and its preparation in order to overcome the defects of poor stability and short shelf life of protein-coupled microspheres in liquid storage in the prior art method, storage method

Method used

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  • Microsphere lyophilized preparation coupled with proteins and preparation method and preservation mode thereof
  • Microsphere lyophilized preparation coupled with proteins and preparation method and preservation mode thereof
  • Microsphere lyophilized preparation coupled with proteins and preparation method and preservation mode thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a microsphere lyophilized preparation coupled with proteins and a preparation method and a preservation mode thereof. The microsphere lyophilized preparation comprises a lyoprotectant and microspheres coupled with the proteins; the lyoprotectant comprises a solvent and a solute; the solvent is phosphate buffer solution with concentration of 0.1 to 0.2mol / L; by volume of thesolvent, the solute comprises 1 to 10% (v / v) of animal serum, 1 to 10% (w / v) of trehalose, 0.2 TO 5% (w / v) of polyvinylpyrrolidone, 0.2 to 5% (w / v) of glucan, 3% to 10% (w / v) of mannitol and 0.01 to0.1% (v / v) of Proclin300; a pH value of the lyoprotectant is 7.4 plus and minus 0.5; the microsphere lyophilized preparation coupled with the proteins is obtained by carrying out lyophilization aftermixing the lyoprotectant and the microspheres coupled with the proteins. By the lyophilized preparation disclosed by the invention, stability of the microspheres coupled with the proteins can be obviously improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a protein-coupled microsphere freeze-dried preparation, a preparation method and a preservation method thereof. Background technique [0002] The main chemical composition of microspheres is polystyrene, and there is a layer of functional base layer (such as amino group, carboxyl group, hydroxyl group, etc.) Microspheres that bind target molecules. [0003] The microspheres of different coding categories are coupled with different ligands, and then the sample to be tested and the reporter molecule are added in sequence. The ligands on different microspheres specifically bind to the target molecule, and the reporter molecule specifically binds to the target molecule. That is to say, a liquid-phase protein chip system is formed, which can simultaneously detect multiple target molecules in the same sample. [0004] The conventional storage method of microspheres is liquid ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/19A61K39/00A61K39/385A61K39/44A61K47/26A61K47/32A61K47/36A61K47/42A61K47/69A61P35/00
CPCA61K9/19A61K47/42A61K47/36A61K47/26A61K47/32A61K47/6927A61K39/44A61K39/385A61K39/001152A61P35/00A61K2039/86
Inventor 赵峰胡海林当黄朝远王增松陈嘉雯赵文博黄璐
Owner GUANGZHOU BIO BLUE TECH CO LTD
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