High carrying capacity protein A immune adsorption material and preparation method thereof

An immunoadsorption material and protein technology, which is applied in the field of high-load protein A immunoadsorption materials and its preparation, can solve the problems of low adsorption performance, low activity, low coupling rate, etc., achieve high reactivity, and improve clinical The effect of the therapeutic effect

Active Publication Date: 2018-03-06
YUNNAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the low activity of the epoxy group and the protein reaction, the coupling rate is low, and the adsorption performance of the prepared protein A immunoadsorbent material on the antibody is also low.

Method used

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  • High carrying capacity protein A immune adsorption material and preparation method thereof
  • High carrying capacity protein A immune adsorption material and preparation method thereof
  • High carrying capacity protein A immune adsorption material and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Preparation of gel microspheres containing primary amino groups

[0039] Take 100mL agarose gel microspheres (Sepharose 6FF), rinse with distilled water, drain, rinse with 20%, 50% and 80% dimethyl sulfoxide in turn, drain, and add to a 1000mL round bottom flask , add 500mL dimethylsulfoxide, 10g sodium hydroxide, 0.3g sodium borohydride, 100mL epichlorohydrin, place in a constant temperature shaker, react at 40°C for 2 hours, then use 80%, 50%, 20 % dimethyl sulfoxide and distilled water to end the reaction. The amount of epoxy groups on the support was detected by the sodium thiosulfate method, and it was measured that there were 100 μmol of epoxy groups per gram of support.

[0040] Put the above product into a 1000mL round bottom flask, add 500ml of 1M ethylenediamine solution (adjust the pH to 9 with hydrochloric acid), react at 40°C for 6 hours, rinse with distilled water, and drain to obtain the primary amino group containing For gel microspheres, it is meas...

Embodiment 2

[0059] 1. Preparation of gel microspheres containing primary amino groups

[0060] Take 100mL of agarose gel microspheres (Sepharose 4FF), rinse with distilled water, drain, add to a 1000mL round bottom flask, add 500mL of 0.2M sodium periodate solution, react at 40°C for 1 hour, Rinse with a large amount of distilled water to end the reaction. According to the concentration of sodium periodate before and after the reaction, it is measured that there are 110 μmol of aldehyde groups per gram of carrier.

[0061] Put the above product into a 1000mL round bottom flask, add 500ml of 0.2M ethylenediamine solution (adjust the pH to 8 with hydrochloric acid), react at 20°C for 4 hours, add 1 gram of sodium borohydride to continue the reaction for 4 hours, Rinse with distilled water and drain to obtain a gel containing primary amino groups. It is measured that there are about 110 μmol of primary amino groups per gram of carrier.

[0062] 2. Preparation of protein A immunosorbent mat...

Embodiment 3

[0069] 1. Preparation of gel microspheres containing primary amino groups

[0070] Take 100mL cellulose microspheres (MT500, 80-100μm), rinse with distilled water, drain, rinse with 20%, 50% and 80% dimethyl sulfoxide in turn, drain, and add to a 1000mL round bottom flask Add 500mL dimethyl sulfoxide, 10g sodium hydroxide, 0.3g sodium borohydride, 100mL epichlorohydrin, place in a constant temperature shaker, react at 20°C for 6 hours, then use 80%, 50%, Wash with 20% dimethyl sulfoxide and a large amount of distilled water to end the reaction. The amount of epoxy groups on the carrier was detected by the sodium thiosulfate method, and it was measured that there were 80 μmol of epoxy groups per gram of carrier.

[0071] Put the above product into a 1000mL round bottom flask, add 500ml of 0.2M ethylenediamine solution (adjust the pH to 11 with hydrochloric acid), react at 20°C for 10 hours, rinse with distilled water, and drain to obtain the primary amino group containing For...

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Abstract

The invention relates to the technical field of blood purification materials and in particular discloses a high carrying capacity protein A immune adsorption material and a preparation method thereof.The protein A immune adsorption material is a high polymer material prepared from hydrophilic gel microspheres and a protein A through coupling; the hydrophilic gel microspheres are adopted as a carrier medium of the material, after being activated with epoxy chloropropane or oxidized with sodium periodate, the carrier medium reacts with ethidene diamine, and then an amino carrier is prepared; the obtained amino carrier reacts with bis-glycidyl ether to generate an activated carrier; and finally the activated carrier is coupled with the protein A. The material is high in content of the protein A in a coupled manner, high in adsorption efficiency on immune globulin and compounds thereof, and is applicable to clinical immune adsorption treatment.

Description

technical field [0001] The invention relates to a blood purification material, in particular to a high-capacity protein A immunoadsorption material and a preparation method thereof. Background technique [0002] Various autoimmune diseases and organ transplant rejection are a series of diseases caused by the production of autoantibodies in the human body against their own tissues and organs or transplanted organs, causing damage to tissues and organs. It is a difficult problem in the treatment of domestic and foreign medical circles one. Autoimmune diseases are caused by the disorder of the human immune system, and have a high disability rate or fatality rate. Common ones include systemic lupus erythematosus (SLE), hemophilia, rheumatoid arthritis (RA), systemic sclerosis (SSc), myasthenia gravis (MG), Sjogren's syndrome (SS), etc. Transplant rejection is the main cause of organ transplant failure, which involves the immune intolerance of the patient's body to the transpla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22B01J20/28B01J20/30C07K1/22
CPCB01J20/22B01J20/28021B01J20/28047B01J2220/4806B01J2220/4812C07K1/22
Inventor 张旭锋王宇杨家梅邓瑶杨海艳郭仁玲
Owner YUNNAN NORMAL UNIV
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