A kind of protein A adsorption material for targeted adsorption and preparation method thereof

An adsorption material and protein technology, which is applied in chemical instruments and methods, suction devices, and other chemical processes, can solve the problems of short coupling spacer arms, high protein A shedding, and low adsorption capacity, so as to improve adsorption performance, High adsorption efficiency and low shedding

Active Publication Date: 2020-04-28
GUANGZHOU KONCEN BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For this reason, we have invented a method of coupling protein A (application number: 201010512308.8, publication number: CN102000550A) using periodate to oxidize and activate agarose. This method uses periodate to oxidize agarose to prepare The agarose gel, and then coupled with protein A, the color of the prepared protein A adsorption material is basically the same as that of the agarose, and the color will not become darker with the use process, but the shedding amount of protein A is higher by this method, And the coupling spacer arm is short, the adsorption capacity is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The synthesis of embodiment 1 epoxy-activated agarose gel

[0044] After washing 100g of agarose gel (Sepharose 6FF) with about 10 times the volume of water for injection, add 100mL of 1M NaOH solution, 30mL of epibromohydrin and 70mL of 1,4-butanediol diglycidyl ether and 0.2 g sodium borohydride, stirred, and reacted at 30° C. for 1.5 h. After the reaction, rinsed with a large amount of water for injection to obtain an epoxy-activated agarose gel.

Embodiment 2

[0045] Synthesis of Example 2 Protein A Adsorption Material

[0046] In a 500mL reaction vessel, add 100mL of epoxy-activated agarose gel synthesized in Example 1, 150mL of borate buffer solution of 0.1mol / L, control the pH value of the system from 7.5 to 8.5, add 14g of genetically engineered recombinant protein A, React in a constant temperature system at 37°C for 20h, stop the reaction, rinse the packing with about 10 times the volume of water for injection, after rinsing, add 200mL of 0.2M ethanolamine solution to block unreacted epoxy groups, and react at 20°C for 10h. After the reaction, rinse with a large amount of water for injection, and finally store it in a solution containing preservatives.

Embodiment 3

[0047] Embodiment 3 adsorption performance contrast

[0048] Take respectively the protein A adsorption material synthesized in Example 2, the protein A adsorption material synthesized using diamine and dialdehyde (the only difference from Example 2) as the spacer arm, and the use of periodate (the only difference from Example 2) ) Each 1mL protein A adsorption material prepared by oxidizing and activating agarose-coupled protein A was installed in a Ф0.8×5cm column, and the column was fully washed with normal saline, and 10mL plasma was passed through the column at a flow rate of 1mL / min. I to G, adsorbed on the adsorption material. Then rinse the plasma with 30 mL of normal saline, use 0.01M citric acid-disodium hydrogen phosphate buffer solution with pH=2.8 as the eluent to elute the human immunoglobulin IgG on the adsorption material, and collect the eluted solution, the eluent was fixed to 100mL, and the absorption peak absorption value of the eluent near 280nm was detec...

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PUM

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Abstract

The invention a protein A adsorption material for targeted adsorption. A protein A is coupled to a high molecular material on a sepharose gel microspherical carrier covalently. The chemical structureof the protein A adsorption material is as follows: a formula as shown in the description represents a sepharose gel, X represents -CH-CN(OH)-CH2- or m is equal to 2,4,6 or n is 2-9, and -NH-SPA is aligand of the protein A. The ligand of the protein A and the sepharose gel microspheres in the adsorption material are connected through ether bonds, the protein A adsorption material is stable in structure, and an adsorbent is stable in color, low in falling amount and high in safety, so that the protein A adsorption material is high in adsorption efficiency and good in regenerability.

Description

technical field [0001] The invention relates to the technical field of medical biomaterials, in particular to a protein A adsorption material for targeted adsorption and a preparation method thereof. Background technique [0002] Immunoadsorption is a new technology developed in the past ten years, which is used to treat some diseases that are difficult to be effective by traditional methods. It combines antigens, antibodies or certain substances with specific physical and chemical affinity as ligands and carriers to make adsorption columns, and uses its specific adsorption properties to selectively or specifically remove endogenous pathogenic factors in patients' blood , so as to achieve the purpose of purifying the blood and relieving the disease. The current clinical application of immunoadsorption can treat a variety of autoimmune diseases and organ transplant rejection by specifically removing autoantibodies, and treat hypercholesterolemia and its complications by adso...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/28B01J20/24B01J20/30A61M1/36
CPCA61M1/3687B01J20/22B01J20/24B01J20/28021B01J20/28047
Inventor 杨正根牛月伟陈校园
Owner GUANGZHOU KONCEN BIOSCI
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