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Synthetic method used for preparing adsorbent for clearing pathogenic antibody by oxidizing periodate

A periodate and synthesis method technology, applied in the field of biomedical materials and blood purification, can solve the problems of harsh elution conditions, antibody instability, unfavorable antibody activity retention, etc., and achieve the effect of simple method process and environmental friendliness

Active Publication Date: 2012-08-29
GUANGZHOU KONCEN BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0008] In addition, in the adsorption media with very promising application prospects, the functional molecules of chimeric proteins (especially Protein LG, Protein LA and Protein AGL) Increased affinity for immunoglobulins leads to harsher antibody elution conditions. For example, 90-95% of immunoglobulins bound to Protein LG can be eluted only at pH 2.0 [Kihlberg, et al.]. It will make the eluted antibody more unstable and not conducive to the retention of antibody activity; at the same time, because the IBP molecule will become unstable under the condition of too low pH, it is easy to degrade and fall off, which is very important for antibody purification production and immune Adsorption therapy is a potential risk factor that cannot be ignored

Method used

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  • Synthetic method used for preparing adsorbent for clearing pathogenic antibody by oxidizing periodate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1) Wash 200 grams of agarose gel (Sepharose 6FF) with about 10 times of water, wash off the ethanol used for preservation, and then mix it with 4 times the mass of 0.05 mol / L sodium periodate solution, and then avoid Under light conditions, react at 30°C for 3 hours, and then wash with a large amount of water to obtain an agarose gel containing aldehyde groups;

[0037] 2) Mix the aldehyde-containing agarose gel with the protein A solution, the amount of the protein A solution is 1 times the mass of the aldehyde-containing agarose gel, in the borate buffer solution with pH 8.4, the protein A solution The concentration was 4 mg / ml, reacted at 30°C for 4 hours, and then washed with a large amount of water to obtain an agarose gel coupled with protein A;

[0038] 3) Add 2 times the pH 7.6 phosphate buffer to the agarose gel coupled with protein A, add 0.001 times the mass of ethanolamine to cap, block at 30°C for 1 hour, and then add 0.001 times in 3 times Quality sodium ...

Embodiment 2

[0041] 1) Wash 200 grams of agarose gel (Sepharose 6FF) with about 10 times of water, wash off the ethanol used for preservation, and then mix it with 0.4 mol / L sodium periodate solution of 0.5 times the mass, and then avoid Under light conditions, react at 40°C for 3 hours, and then wash with a large amount of water to obtain an agarose gel containing aldehyde groups;

[0042] 2) Mix the aldehyde-containing agarose gel with the protein A solution, the amount of the protein A solution is twice the mass of the aldehyde-containing agarose gel, and in the carbonate buffer solution of pH 11, the protein A solution The concentration was 3 mg / ml, reacted at 30°C for 4 hours, and then washed with a large amount of water to obtain an agarose gel coupled with protein A;

[0043] 3) Add 2 times the pH 7.6 phosphate buffer to the agarose gel coupled with protein A, add 0.5 times the mass of ethanolamine to cap, block at 30°C for 1 hour, and then add 0.001 times in 3 times Quality sodium...

Embodiment 3

[0046] 1) Wash 200 grams of agarose gel (Sepharose 6FF) with about 10 times of water, wash off the ethanol used for preservation, and then mix it with 0.2 mol / L potassium periodate solution of 2 times the mass, and then avoid Under light conditions, react at 30°C for 5 hours, and then wash with a large amount of water to obtain an agarose gel containing aldehyde groups;

[0047] 2) Mix the aldehyde-containing agarose gel with the protein A solution, the amount of the protein A solution is 4 times the mass of the aldehyde-containing agarose gel in the acetate buffer of pH 4, the concentration is 2 mg / ml , reacted at 40°C for 8 hours, and then washed with a large amount of water to obtain an agarose gel coupled with protein A;

[0048] 3) Add protein A-coupled agarose gel to 2 times the pH 7.6 phosphate buffer, add 0.001 times the mass of glycine ethyl ester to cap, block at 30°C for 1 hour, and then add in 3 times 0.02 times the mass of sodium borohydride is reduced for 24 hou...

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Abstract

The invention relates to a synthetic method used for preparing an adsorbent for clearing a pathogenic antibody by oxidizing periodate, and discloses a method for preparing a protein immunoadsorption material by oxidizing the periodate by taking agarose gel as a carrier. The method comprises the following steps of: preparing aldehyde group-containing agarose gel by oxidizing the agarose gel by using the periodate; coupling the aldehyde group-containing agarose gel and immunoglobulin binding protein; and blocking a material and reducing or reducing directly without blocking to obtain the immunoadsorption blood purification material. The method has the characteristics of simple process, environmental friendliness, low cost and the like; and simultaneously the product has high specificity, adsorptive property, regenerability and the like, and can be used for clinical immunoadsorption treatment practically.

Description

technical field [0001] The invention relates to a synthesis method for preparing an adsorbent for eliminating pathogenic antibodies by oxidation of periodate, and belongs to the fields of biomedical materials and blood purification. Background technique [0002] Immunoadsorption (IA) therapy is a new blood purification technology developed in the past 15 years, and it is used to treat some diseases that traditional methods are ineffective. The basic principle is to connect antigens, antibodies or some substances with specific affinity as ligands to the carrier, prepare an in vitro adsorption column for in vitro blood purification, and use its specific adsorption properties to selectively or specifically remove Endogenous pathogenic factors in the patient's blood, so as to achieve the purpose of purifying the blood and relieving the disease. In 2001, the first European Immunoadsorption Symposium was held in London, England. More than 200 experts and scholars from 17 countrie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/24B01J20/30
Inventor 陈校园张旭锋许春生艾福金余波光
Owner GUANGZHOU KONCEN BIOSCI
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