An affinity biomimetic chromatography medium with tetrapeptide as functional ligand
A chromatographic medium and ligand technology, applied in the field of protein chromatographic separation technology, can solve the problems of high price of protein A affinity medium, harsh elution conditions, easy detachment of ligands, etc., to avoid antibody aggregation or activity decline, The effect of large adsorption capacity and strong adsorption selectivity
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Embodiment 1
[0042] Example 1: Preparation of Tetrapeptide Affinity Chromatography Medium
[0043] The process of preparing tetrapeptide chromatography medium with agarose gel as matrix mainly includes four steps: matrix activation, bromoalcoholation, spacer coupling and ligand coupling. (1) Matrix activation: first weigh 5g of sodium hydroxide, add 10mL of 20% (v / v) DMSO solution, dissolve, let stand, after the solution is cooled, add 10g of drained agarose gel matrix, and then add 10mL Allyl bromide, activated in a shaker at 180rpm at 30°C for 24 hours, washed with deionized water, and filtered with suction to obtain an activated matrix; (2) bromoalcoholization: mix the activated matrix with 5g NBS, and react in a shaker at 180rpm at 30°C For 3 hours, wash with deionized water and filter with suction to obtain the brominated matrix; (3) space arm coupling: mix the brominated matrix with 3 mL of hexamethylenediamine and 1M sodium carbonate buffer (pH 12), at 30°C at 180rpm React in a sha...
Embodiment 2
[0044] Example 2: Preparation of Tetrapeptide Affinity Chromatography Medium
[0045] The process of preparing tetrapeptide chromatography medium with agarose gel as matrix mainly includes four steps: matrix activation, bromoalcoholation, spacer coupling and ligand coupling. (1) Matrix activation: first weigh 5g of sodium hydroxide, add 10mL of 20% (v / v) DMSO solution, dissolve, let stand, after the solution is cooled, add 10g of drained agarose gel matrix, and then add 10mL Allyl bromide, activated in a shaker at 180rpm at 30°C for 24 hours, washed with deionized water, and filtered with suction to obtain an activated matrix; (2) bromoalcoholization: mix the activated matrix with 5g NBS, and react in a shaker at 180rpm at 30°C For 3 hours, wash with deionized water and filter with suction to obtain the brominated matrix; (3) space arm coupling: mix the brominated matrix with 3 mL of hexamethylenediamine and 1M sodium carbonate buffer (pH 12), at 30°C at 180rpm React in a sha...
Embodiment 3
[0046] Example 3: Preparation of Tetrapeptide Affinity Chromatography Medium
[0047] The process of preparing tetrapeptide chromatography medium with agarose gel as matrix mainly includes four steps: matrix activation, bromoalcoholation, spacer coupling and ligand coupling. (1) Matrix activation: first weigh 5g of sodium hydroxide, add 10mL of 20% (v / v) DMSO solution, dissolve, let stand, after the solution is cooled, add 10g of drained agarose gel matrix, and then add 10mL Allyl bromide, activated in a shaker at 180rpm at 30°C for 24 hours, washed with deionized water, and filtered with suction to obtain an activated matrix; (2) bromoalcoholization: mix the activated matrix with 5g NBS, and react in a shaker at 180rpm at 30°C For 3 hours, wash with deionized water and filter with suction to obtain the brominated matrix; (3) space arm coupling: mix the brominated matrix with 3 mL of hexamethylenediamine and 1M sodium carbonate buffer (pH 12), at 30°C at 180rpm React in a sha...
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