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Biomimetic specific immune adsorption material with PAMAM (Polyamidoamine) as spacer arm, and preparation method and application thereof

An immunoadsorption material and biomimetic technology, which is applied in the field of preparation of biomimetic specific immunoadsorption materials for blood purification, can solve the problems of protein difficult to sterilize, cannot be coupled to the carrier, and reduce the effect of immunoadsorption, etc., to achieve pro- Good water and blood compatibility, good chemical stability and mechanical strength, avoid expensive effects

Active Publication Date: 2013-10-30
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, protein-based ligands have some defects that cannot be ignored: proteins are difficult to withstand the harsh disinfection process and are easy to fall off from the carrier, and these fallen-off ligands will cause safety problems; the high price of protein-based immunoadsorbent materials has caused great harm to patients. Heavy economic burden; protein ligands are difficult to modify with chemical functional groups, so they cannot be coupled to the carrier at the most suitable angle
The spacer arm commonly used in immunoadsorption materials is a linear spacer arm, and this type of spacer arm will entangle with biomacromolecules when separating biomacromolecules, thereby reducing the effect of immunoadsorption

Method used

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  • Biomimetic specific immune adsorption material with PAMAM (Polyamidoamine) as spacer arm, and preparation method and application thereof
  • Biomimetic specific immune adsorption material with PAMAM (Polyamidoamine) as spacer arm, and preparation method and application thereof
  • Biomimetic specific immune adsorption material with PAMAM (Polyamidoamine) as spacer arm, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Preparation of Azide Sepharose

[0060] In a 100 mL conical flask, add 4.0 g of drained agarose gel, 8 mL of epichlorohydrin and 4 mL of 3M sodium hydroxide solution. After the mixture was stirred and reacted at room temperature for 3 hours, washed with water and drained until the phenolphthalein did not turn red after adding 1.3M sodium thiosulfate to the washing solution to obtain epoxidized agarose. Dissolve 1.82 g of sodium azide in 15 mL of water, add 4.0 g of epoxidized agarose, and react at 20 °C for 60 h. After the reaction is completed, wash and drain to obtain azide agarose gel. attached figure 1 Infrared spectrum shown at 2100cm -1 The characteristic absorption peak of azide group appeared at azide agarose, which proved the existence of azide group in azide agarose.

[0061] Synthesis of G3.0PAMAM Dendrimers

[0062] In a 250ml round-bottomed flask, first add 200ml of methanol, then add 272mmol of methyl acrylate, put it in an ice-water bath for a period...

Embodiment 2

[0078] Preparation of Azide Sepharose

[0079] In a 100mL Erlenmeyer flask, add 4.0g of drained agarose gel, 8mL of epichlorohydrin and 8mL of 1.5M sodium hydroxide solution. The mixture was stirred and reacted at room temperature for 4 hours, washed with water and drained until the phenolphthalein did not turn red after adding 1.3M sodium thiosulfate to the washing solution to obtain epoxidized agarose. Dissolve 0.78 g of sodium azide in 10 mL of water, add 4.0 g of epoxidized agarose, and react at 40 °C for 30 h. After the reaction is completed, wash and drain to obtain azide agarose gel.

[0080] Synthesis of G2.0PAMAM Dendrimers

[0081] In a 250ml round-bottomed flask, first add 270ml of methanol, then add 272mmol of methyl acrylate, place in an ice-water bath for a period of time to make the temperature of the system about 10°C, and then continuously dropwise add 54.4mmol of propargylamine within 1h. 30 ml of methanol solution was added dropwise, and the reaction was ...

Embodiment 3

[0096] Preparation of Azide Sepharose

[0097] In a 100mL Erlenmeyer flask, add 4.0g of drained agarose gel, 8mL of epichlorohydrin and 6mL of 2.5M sodium hydroxide solution. The mixture was stirred and reacted at room temperature for 3.5 h, washed with water and drained until the phenolphthalein did not turn red after adding 1.3 M sodium thiosulfate to the washing solution, thus obtaining epoxidized agarose. Dissolve 3.9 g of sodium azide in 25 mL of water, add 4.0 g of epoxidized agarose, and react at 60 °C for 6 h. After the reaction is completed, wash and drain to obtain azide agarose gel.

[0098] Synthesis of G4.0PAMAM Dendrimers

[0099] In a 250ml round-bottomed flask, first add 180ml of methanol, then add 300mmol of methyl acrylate, put it in an ice-water bath for a period of time to make the temperature of the system about -10°C, and then continuously dropwise add 20.0mmol of propargylamine within 1h 30ml of methanol solution was added dropwise, and the reaction w...

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Abstract

The invention discloses a biomimetic specific immune adsorption material with PAMAM (Polyamidoamine) as a spacer arm, and a preparation method and the application thereof. The preparation process comprises the following steps of: firstly, obtaining sepharose gel azide by reaction of agarose epoxide and sodium azide; preparing a series of PAMAM dendrimer-like polymers in different generations by repeated addition reaction with methyl acrylate and amination reaction with ethidene diamine; after preparing amino acid methyl ester hydrochloride with the amino acid as the raw material, modifying the PAMAM dendrimer-like polymers with the amino acid methyl ester hydrochloride to prepare the amino acid modified PAMAM; coupling the amino acid modified PAMAM with sepharose gel azide through a click reaction so as to obtain the biomimetic specific immune adsorption material with PAMAM as the spacer arm. The material disclosed in the invention is prepared by simple synthetic steps, and the preparation is safe; products have the characteristics of strong specificity, high adsorbing efficiency to the immunoglobulin and excellent regeneration performance, and can be used for IgG (immunoglobulin G) separation and clinical immune adsorption treatment.

Description

technical field [0001] The invention belongs to biomedical equipment, and particularly provides a method for preparing a biomimetic specific immunoadsorption material for blood purification. The adsorption material uses a polyamide-amine dendrimer (PAMAM) as a spacer arm. technical background [0002] It is well known that protein-based immunoadsorbent materials with protein A, G, etc. as ligands have good biospecific affinity for immunoglobulin (IgG). The interaction between protein ligands and IgG involves hydrophobic interaction and electrostatic interaction. Immunoadsorbent materials containing such biospecific ligands have been proven to remove autoantibodies to purify antibodies and treat various autoimmune diseases, so they have been widely used in biological purification and clinical applications. However, protein-based ligands have some disadvantages that cannot be ignored: proteins are difficult to withstand harsh sterilization processes and are easily detached fro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K17/02B01J20/26B01J20/30B01D15/08C07K16/00C07K1/22
Inventor 李光吉胡小艳
Owner SOUTH CHINA UNIV OF TECH
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