Blood purifying protein A immunoadsorption material and synthesizing method thereof
An immunoadsorption material and blood purification technology, which is applied in the field of protein A immunoadsorption material and its synthesis for blood purification treatment, can solve the problems of large batch-to-batch quality differences, increased industrialization costs, and unstable performance of adsorption materials, and achieve Improve the clinical treatment effect, improve the binding efficiency, and have good regenerative performance
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Embodiment 1
[0037] Synthesis of Active Sepharose 6FF Agarose Gel Containing Epoxy Groups
[0038] Add 1 liter of Sepharose 6FF agarose gel, 1500 milliliters of 2.5 mol / L NaOH aqueous solution, 3 grams of sodium borohydride in a 5-liter 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
[0040] Synthesis of active Sepharose 6FF agarose gel containing amino groups
[0041] 1. Reaction of epoxy-active Sepharose 6FF agarose gel with diethylenetriamine
[0042] 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 200mL of diethylenetriamine and react at a constant temperature of 50°C for 2 hours. After the reaction stopped, rinse with 1mol / L sodium chloride solution and sterile water to remove residual diethylenetriamine to obtain active Sepharose 6FF agarose gel containing amino groups.
[0043] 2. Reaction of epoxy-active Sepharose 6FF agarose gel with ethylenediamine
[0044] 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 r...
Embodiment 3
[0048] Synthesis of Sepharose 6FF Agarose Gel Containing Carbonylimidazole
[0049] 1 liter of amino-active Sepharose 6FF agarose gel synthesized in "Example 2" (1, 2 or 3) was successively treated with 25%, 50%, 75%, 100% (v / v) of anhydrous dioxane The ring gradient solution was rinsed, and the gel was gradually transferred from the aqueous phase to anhydrous dioxane solvent, and then added to a 5L reactor, followed by 1500ml of anhydrous dioxane solvent, 60 g of N,N'- Carbonyldiimidazole was reacted at a temperature of 20°C for 2 hours. After stopping the reaction, rinse with 100%, 75%, 50%, 25% (v / v) dioxane gradient concentrated solution and distilled water at 4°C successively, and gradually transfer the carrier to the water phase to obtain carbonyl imidazolyl Sepharose 6FF agarose gel, proceed to the next reaction immediately.
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