Chromatographic medium and preparation method thereof
A chromatographic medium and matrix technology, applied in the field of chromatographic medium and its preparation, can solve the problems that the purification efficiency has not been significantly improved, cannot remove protein A well, and has limited adsorption performance, so as to achieve continuous separation and adsorption characteristics, The effect of low catalyst and ligand usage and high ligand coupling efficiency
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[0038] Preparation Example 1
[0039] The chromatography medium is prepared by the following steps:
[0040] Step 1) Wash 10g of agarose gel microspheres with deionized water, drain it, add 0.015g of sodium borohydride and 0.5g of anhydrous sodium sulfate, and then add 1.5mL of 50% sodium hydroxide solution, 50% React at °C for 1 hour. Then add 3.5mL allyl glycidyl ether, activate at 50℃ for 8 hours, filter with suction, wash with 50ml deionized water, 50ml absolute ethanol, 50ml 0.2M acetic acid and 100ml deionized water to obtain activated chromatography Matrix
[0041] Step 2). Mix 10g of activated chromatography matrix, 0.1g of sodium acetate and 1.5ml of deionized water to obtain a mixed solution. While stirring the mixed solution at room temperature, add bromine water dropwise until the mixed solution turns pale yellow and The color does not change within 1 minute, then add sodium formate dropwise until it becomes colorless, and finally filter and clean to obtain a brominate...
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[0043] Preparation Examples 2-7
[0044] Preparation Examples 2-7 were prepared according to the method of Example 1, except that the amount of reactants and reaction time in Example 2-7 were slightly different from those in Example 1. The prepared chromatographic media The ligand density is different. See Table 1 below for details.
[0045]
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[0046] Example 1
[0047] Using the chromatography medium Adhere NUPharose FF prepared in Preparation Example 6, the saturated adsorption capacity Q for bovine IgG was measured at 25°C and different pH. m , Determine its adsorption isotherm, the results are shown in figure 1 in.
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