Affinity chromatography filler dynamic loading capacity determination method

2016 
The invention discloses an affinity chromatography filler dynamic loading capacity determination method.The method comprises the steps that firstly, purified protein samples with different concentrations are prepared with flowthrough liquid; secondly, a filtering plate for filling of filler is prepared; thirdly, the samples are added onto the filtering plate and hatched; fourthly, suction filtration is carried out, the flowthrough liquid is collected, the protein concentrations are detected, and an adsorption isotherm is fitted; fifthly, chromatographic columns are filled and loaded with the samples, elution is carried out, the retention volume of ultraviolet outlet peaks and the retention volume of electric conductance outlet peaks are recorded, the chromatographic columns are short circuited, the retention volume of electric conductance outlet peaks is recorded, and the water drainage porosity outside the filler, the full-permeation porosity and the filler porosity are calculated; sixthly, the average radius of protein hydration molecules is determined; seventhly, the dynamic loading capacity is calculated.According to the affinity chromatography filler dynamic loading capacity determination method, according to the characteristics and natures of the affinity filler and monoclonal antibodies, an adsorption isotherm and adsorption dynamical model is studied through the high throughput screening technology, the dynamic loading capacity of the chromatography filler is predicted through the static loading capacity of the monoclonal antibodies, less time is consumed, cost is low, and the accuracy of a determination result is high.
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