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Protein A immunoadsorption material for eliminating pathogenic antibody and its complexes, and synthesizing method and application thereof

A technology of immunoadsorbent materials and complexes, which is used in the field of protein A immunoadsorbent materials for removing pathogenic antibodies and their complexes and its synthesis and application, and can solve the problem of insufficient length of spacer arms and low ammonia reactivity of immunoadsorbent materials , Low antibody adsorption efficiency and other issues, to achieve the effect of improving clinical treatment effect, improving binding efficiency, and good regeneration performance

Active Publication Date: 2010-05-26
GUANGZHOU KONCEN BIOSCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the small size of the ammonia water molecule, the spacer arm of the obtained immunoadsorbent material is not long enough. In addition, we found in the experiment that the ammonia water reaction activity is low, the coupling protein A is less, and the protein A immunoadsorbent material obtained by synthesis has a low adsorption efficiency for antibodies.

Method used

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  • Protein A immunoadsorption material for eliminating pathogenic antibody and its complexes, and synthesizing method and application thereof
  • Protein A immunoadsorption material for eliminating pathogenic antibody and its complexes, and synthesizing method and application thereof
  • Protein A immunoadsorption material for eliminating pathogenic antibody and its complexes, and synthesizing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Synthesis of Active Sepharose 6FF Agarose Gel Containing Epoxy Groups

[0042] Add 1 liter of trade name Sepharose 6FF agarose gel, 1500 ml of 2.5 mol / L NaOH aqueous solution, 3 grams of sodium borohydride into a 5 L reactor, mix well, add about 300 mL of epibromohydrin, and place In a constant temperature shaker, react at 30°C for 4 hours. After the reaction was completed, the gel was filtered and rinsed with a large amount of distilled water until neutral. Store the activated medium at 4°C for later use. The number of epoxy groups in the gel activated by this method was detected by the sodium thiosulfate method, and it was measured that there were at least 50 μmol of epoxy active groups per milliliter.

Embodiment 2

[0044] Synthesis of active Sepharose 6FF agarose gel containing amino groups

[0045] 1. Reaction of epoxy-active Sepharose 6FF agarose gel with diethylenetriamine

[0046] Add 1 liter of epoxy-active Sepharose 6FF agarose gel synthesized in Example 1, 1.5 L of 0.1 mol / L borate buffer solution, and control the pH value in the range of 7.0 to 9.0. Ethylenetriamine was reacted at a constant temperature of 50°C for 2 hours. After the reaction stopped, rinse with 1mol / L sodium chloride solution and disinfected water in large quantities to remove residual diethylenetriamine to obtain an activated agarose gel containing amino groups.

[0047] 2. Reaction of epoxy-active Sepharose 6FF agarose gel with ethylenediamine

[0048] In the 5L reactor, add 1 liter of epoxy-active Sepharose 6FF agarose gel synthesized in "Example 1", 1.5L of borate buffer solution of 0.1mol / L, and the pH value is controlled in the scope of 7.0~9.0, Add 160 mL of ethylenediamine, and react at a constant tem...

Embodiment 3

[0052] Synthesis of Active Sepharose 6FF Agarose Gel Containing Aldehyde Group

[0053] Add 1 liter of aminoactive Sepharose 6FF agarose gel synthesized in "embodiment two" (1, 2 or 3) in the reactor of 5L, 0.1mol / L borate buffer solution 1.2mL, control system pH value at 8 or so, add 200mL glutaraldehyde, and react at a constant temperature of 30°C for 3 hours. After the reaction was stopped, the residual glutaraldehyde was washed away with a large amount of water to obtain an agarose gel containing aldehyde groups.

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Abstract

The invention provides a protein A immunoadsorption material which eliminates pathogenic antibody and the compound from plasma and the preparation method thereof. The invention discloses a polymer material coupled by agarose gel and staphylococcal protein A (SPA) and the material takes the agarose gel as vector matrix and takes polyamines reagent as a space arm, thus reacting with glutaraldehyde and finally coupling with the protein A and blocking, reducing a double bond after being activated by epoxy bromopropane. The product has high safety; by making use of the produce, the pathogenic antibody, more particularly, the immune compound pathogenic antibody can be effectively and selectively eliminated from the plasma of patients; and the invention can be applied to clinical immunoadsorptiontreatments.

Description

Technical field: [0001] The invention relates to a protein A immunoadsorbent material for blood purification treatment and a preparation method thereof, which can selectively remove pathogenic antibodies in patients' plasma, especially the pathogenic antibodies in the form of immune complexes. 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 some substances with specific physical and chemical affinity as ligands and carriers to make adsorption columns, and uses its high specific adsorption performance to selectively or specifically remove pathogenic factors in the patient's blood, thereby To achieve the purpose of purifying blood and relieving symptoms. The current clinical application of immunoadsorption can treat autoimmune diseases and organ transplant rejection by specifically removing auto...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/22
Inventor 陈校园张旭锋汪志刚李光吉许春生
Owner GUANGZHOU KONCEN BIOSCI
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