CN110624422A

CN110624422A - Dialysis membrane for removing carbonylation protein and preparation method thereof

Info

Publication number: CN110624422A
Application number: CN201910762285.5A
Authority: CN
Inventors: 高明; 郑立; 杨屹峰
Original assignee: Guangxi Medical University
Current assignee: Guangxi Medical University
Priority date: 2019-08-19
Filing date: 2019-08-19
Publication date: 2019-12-31

Abstract

The invention relates to a dialysis membrane for removing carbonylation protein, which is obtained by performing hydrazide group modification on a cellulose membrane, and also provides a preparation method of the dialysis membrane for removing the carbonylation protein, wherein the cellulose membrane is soaked in dimethyl sulfoxide, activated by 1, 1' -carbonyldiimidazole and then reacted with hydrazine. The hydrazide group combined on the cellulose membrane can be specifically combined with the carbonylation protein, so the cellulose membrane modified by the hydrazide group can effectively remove the carbonylation protein without removing the normal protein, and the preparation method is simple and efficient.

Description

Dialysis membrane for removing carbonylation protein and preparation method thereof

Technical Field

The invention relates to the technical field of dialysis membranes, in particular to a dialysis membrane for removing carbonylation proteins and a preparation method thereof.

Background

Hemodialysis is a commonly used method of kidney replacement therapy in clinical practice for patients with end-stage renal disease, and in short, hemodialysis is a process of removing toxic substances from blood by using a dialysis membrane to maintain homeostasis. In previous researches, a conventional low-flux hemodialysis membrane can remove small molecular toxins in a body, but cannot effectively remove medium and large molecular toxins in blood; high-throughput hemodialysis membranes are effective in removing large toxic substances such as carbonylated proteins from blood, but at the same time, some normal, healthy large proteins are removed. It is known that the carbonylation protein is an important toxin protein produced during the development of end stage renal disease, and therefore, there is a need to develop a dialysis membrane that can selectively remove the carbonylation protein but not the normal protein.

Disclosure of Invention

The invention provides a dialysis membrane capable of removing carbonylation protein but not removing normal protein to solve the technical problems.

The technical scheme for solving the technical problems is as follows: a dialysis membrane for removing carbonylated proteins, which is obtained by subjecting a cellulose membrane to hydrazide-based modification.

The invention has the beneficial effects that: the hydrazide group combined on the cellulose membrane can be combined with the carbonylation protein, and the cellulose membrane modified by the hydrazide group can effectively remove the carbonylation protein without removing normal protein.

The invention also provides a preparation method of the dialysis membrane for removing the carbonylation protein, which is prepared by immersing a cellulose membrane in dimethyl sulfoxide, activating by 1, 1' -carbonyldiimidazole and then reacting with hydrazine.

Further, the hydrazine is hydrazine hydrate.

Further, the method comprises the following specific steps: soaking a cellulose membrane in dimethyl sulfoxide, adding 1, 1' -carbonyldiimidazole to react for 6-8 h, then dropwise adding a hydrazine hydrate solution, continuing to react for 12-24 h, and then washing with deionized water to obtain the dialysis membrane for removing the carbonylated protein.

Furthermore, 1-3 mL of dimethyl sulfoxide, 0.1-0.3 g of 1, 1' -carbonyldiimidazole, 0.2-0.8 mL of hydrazine hydrate and 80% of hydrazine hydrate are added to each cellulose membrane with the diameter of 25 mm.

Further, the pore size of the cellulose membrane was 0.45 μm.

The method has the advantages of simple and efficient preparation method, high hydrazide modification level on the prepared dialysis membrane and high removal efficiency of the carbonylation protein.

Detailed Description

The principles and features of this invention are described below in conjunction with specific embodiments, which are set forth merely to illustrate the invention and are not intended to limit the scope of the invention.

The materials and drugs used in the present invention include cellulose membranes (25 mm in diameter, 0.45 μm in pore size, VWR corporation), dimethylsulfoxide (DMSO, Sigma, 99.9%) 1, 1' -carbonyldiimidazole (CDI, 97.0% or more, Sigma) and hydrazine hydrate (78-82%, Sigma).

Example 1

Soaking a piece of commercial Cellulose Membrane (CM) in 1mL of dimethyl sulfoxide, adding 0.2g of 1, 1' -carbonyldiimidazole for activation for 6h, dropwise adding 0.4mL of hydrazine hydrate solution, continuing to react for 12h, and finally washing the cellulose membrane with a large amount of deionized water to obtain the dialysis membrane for removing the carbonylated proteins.

Example 2

Soaking a piece of commercial Cellulose Membrane (CM) in 3mL of dimethyl sulfoxide, adding 0.3g of 1, 1' -carbonyldiimidazole for activation for 7h, dropwise adding 0.8mL of hydrazine hydrate solution, continuing to react for 15h, and finally washing the cellulose membrane with a large amount of deionized water to obtain the dialysis membrane for removing the carbonylated proteins.

Example 3

Soaking a piece of commercial Cellulose Membrane (CM) in 2mL of dimethyl sulfoxide, adding 0.1g of 1, 1' -carbonyldiimidazole for activation for 8h, dropwise adding 0.2mL of hydrazine hydrate solution, continuing to react for 24h, and finally washing the cellulose membrane with a large amount of deionized water to obtain the dialysis membrane for removing the carbonylated proteins.

Verification example 1

Mixing 8-16ml of 0.5-1mg/ml bovine serum albumin with 500-1000 mu mol of acrolein, standing overnight in the dark to obtain carbonylated bovine serum albumin, measuring absorbance at 370nm by ultraviolet-visible spectroscopy, and calculating the content of carbonylated protein according to a standard curve. The amount of carbonylated bovine serum albumin in the phosphate buffer was determined to be 8 mg. The dialysis membranes for removing the carbonylation proteins obtained in the embodiments 1 to 3 are respectively soaked in the carbonylation bovine serum albumin solution for 12 to 24 hours, the content of the carbonylation proteins in the solution is measured and calculated again, the amount of the carbonylation bovine serum albumin is respectively reduced to 4.8mg, 4.2mg and 3.9mg after the treatment of the dialysis membranes of the embodiments 1 to 3, and the removal efficiency of the carbonylation bovine serum albumin is respectively 40%, 47.5% and 51.25%.

Verification example 2

A certain amount of serum of a patient with end-stage renal disease is taken, diluted by a certain amount of phosphate buffer solution, and the content of the carbonylation protein is calculated to be 12 g by measuring the absorbance at the wavelength of 370nm through an ultraviolet-visible spectrum method. The dialysis membranes for removing carbonylated proteins obtained in examples 1 to 3 were immersed in the serum diluent of a patient, and the content of carbonylated proteins in the diluent was measured and calculated again, and after the treatment with the dialysis membranes in examples 1 to 3, the content of carbonylated proteins in the serum diluent of the patient was reduced to 5.4mg, 6.2mg and 6.6mg, respectively, and the removal efficiency of carbonylated proteins in the serum of the patient was 55%, 48.3% and 45%, respectively.

The dialysis membrane for removing the carbonylation protein is a low-flux dialysis membrane, can remove redundant water and small-molecule toxin in blood, and simultaneously, the hydrazide group combined with the dialysis membrane can be specifically combined with the aldehyde group or the ketone group on the carbonylation protein, thereby selectively removing the carbonylation protein without losing normal protein.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims

1. A dialysis membrane for removing a carbonylated protein, which is obtained by subjecting a cellulose membrane to hydrazide-based modification.

2. A method of preparing a dialysis membrane for the removal of carbonylated proteins as claimed in claim 1, wherein said membrane is prepared by immersing a cellulose membrane in dimethyl sulfoxide, activating with 1, 1' -carbonyldiimidazole and reacting with hydrazine.

3. The method according to claim 2, wherein the hydrazine is hydrazine hydrate.

4. The preparation method according to claim 3, comprising the following steps: soaking a cellulose membrane in dimethyl sulfoxide, adding 1, 1' -carbonyldiimidazole to react for 6-8 h, then dropwise adding a hydrazine hydrate solution, continuing to react for 12-24 h, and then cleaning with deionized water to obtain the dialysis membrane for removing the carbonylation protein.

5. The method according to claim 4, wherein 1 to 3mL of dimethyl sulfoxide, 0.1 to 0.3g of 1, 1' -carbonyldiimidazole, 0.2 to 0.8mL of hydrazine hydrate, and the concentration of hydrazine hydrate is 80% are added to each cellulose film having a diameter of 25 mm.

6. The production method according to claim 5, wherein the pore diameter of the cellulose membrane is 0.45 μm.