Synthetic method used for preparing adsorbent for clearing pathogenic antibody by oxidizing periodate
A periodate and synthesis method technology, applied in the fields of biomedical materials and blood purification, can solve the problems of harsh elution conditions, unstable antibodies, easy degradation and shedding, etc., and achieve the effect of environmental friendliness and simple method process
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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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