Biomacromolecule microreactor based on protein film isolation and fixation

A biomacromolecule and microreactor technology, applied in the field of biomacromolecule microreactors, can solve problems such as poor biocompatibility, high cost, and complicated preparation methods, and achieve easy operation, low cost, and simple and efficient preparation process Effect

Inactive Publication Date: 2019-07-19
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of its preparation methods are complicated, costly, and poor in biocompatibility, such as layer-by-layer self-assembly technology (LBL), which is difficult to meet the urgent needs of industry.

Method used

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  • Biomacromolecule microreactor based on protein film isolation and fixation
  • Biomacromolecule microreactor based on protein film isolation and fixation
  • Biomacromolecule microreactor based on protein film isolation and fixation

Examples

Experimental program
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Effect test

Embodiment 1

[0036] 1. Prepare 5mL 0.33mol / L Na respectively 2 CO 3 Aqueous solution and 0.33mol / L CaCl 2 aqueous solution, and then 100 μL of 10 mg / mL fluorescently labeled bovine serum albumin aqueous solution was added to the CaCl 2 solution in aqueous solution, then add Na 2 CO 3 Aqueous solution, and kept stirring at 650rpm for 30 seconds, after standing for 5 minutes, centrifuged and washed with deionized water to obtain CaCO with bovine serum albumin loaded inside and on the surface 3 particles. Such as Figure 1~6As shown, CaCO with bovine serum albumin loaded inside and on the surface was successfully prepared 3 particles.

[0037] 2. Adjust 1mL of 10mmol / L tris(2-carboxyethyl)phosphine in PBS buffer to pH 7.0 with NaOH, then mix it with 1mL of 2mg / mL lysozyme in PBS buffer, and add the above preparation Well loaded CaCO with bovine serum albumin 3 Particles were incubated at room temperature for 50 minutes; then the reaction impurities were removed by centrifugation to o...

Embodiment 2

[0040] In step 1 of this embodiment, prepare 5mL 0.33mol / L Na 2 CO 3 Aqueous solution and 0.33mol / L CaCl 2 aqueous solution, then Na 2 CO 3 Aqueous solution was added to CaCl 2 in aqueous solution, and kept stirring at 650rpm for 30 seconds, after standing still for 5 minutes, it was washed by centrifugation with deionized water to obtain CaCO 3 particles. Will get CaCO 3 The particles were dispersed in 100 μL of 10 mg / mL fluorescently labeled bovine serum albumin aqueous solution, and incubated at room temperature for 24 hours to obtain CaCO with bovine serum albumin loaded on the surface. 3 particles, such as Figures 12 to 13 shown. The other steps were the same as in Example 1 to obtain a bovine serum albumin microreactor isolated and fixed by a lysozyme film.

Embodiment 3

[0042] In step 1 of this embodiment, prepare 5mL 0.33mol / L Na 2 CO 3 Aqueous solution and 0.33mol / L CaCl 2 aqueous solution, and then 100 μL of 10 mg / mL fluorescently labeled bovine serum albumin aqueous solution and 100 μL of 10 mg / mL fluorescently labeled human serum albumin aqueous solution were added to CaCl 2 In the solution aqueous solution, other steps are identical with embodiment 1, obtain the microreactor (see Figure 14 ).

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Abstract

The invention discloses a biomacromolecule microreactor based on protein film isolation and fixation. Firstly, functional biomacromolecules needing isolation are loaded in the inner parts and the surfaces of CaCO3 granules by a co-precipitation method, or are loaded on the surfaces of the CaCO3 granules by a direct adsorption method, then a protein film is used for wrapping the surfaces of the CaCO3 granules, and finally, an internal CaCO3 formwork is removed through EDTA or etching with an acid solution, so that isolation and fixation of the functional biomacromolecules on the microcosmic level are realized, and further the development of physiological functions of the biomacromolecules is not influenced. The microreactor is simple in preparation method, low in cost and environmentally-friendly, microcosmic isolation of the functional biomacromolecules including various protein, enzymes, antibodies, lipid, polypeptide, DNA, RNA and the like can be realized, the microreactor has broaduniversality, large-scale industrial application is easy to realize, and the size and the shape of the microreactor for performing isolation on the biological macromolecules can be adjusted and controlled through the formwork.

Description

technical field [0001] The invention relates to a biomacromolecule micro-reactor which realizes the isolation and fixation of various functional biomacromolecules at the microscopic level based on the amyloid protein film, and then does not affect each other to exert their respective physiological functions. Background technique [0002] In organisms, the vast majority of material conversions are catalyzed by many kinds of enzymes in series. In order to ensure the high efficiency of tandem catalysis, these cooperating enzymes are usually concentrated in certain cells or organelles. This physiological structure can not only effectively reduce the loss of intermediate products, but also prevent the toxic effects of by-products in the process of material conversion on the body. Therefore, it is very important for the encapsulation and fixation of biological macromolecules. Metal, inorganic or protein three-dimensional capsules or microreactors with special colloidal structure...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K17/14C12N11/14C12N15/10C07K16/00
CPCC07K17/14C12N11/14C12N15/10C07K16/00
Inventor 杨鹏刘瑞瑞
Owner SHAANXI NORMAL UNIV
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