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Determination method for dynamic loading capacity of affinity chromatography filler

A technology of chromatography filler and determination method, which is applied in chemical instruments and methods, analytical materials, material separation, etc., can solve the problems of large material consumption, long process time, and long processing cycle, and achieve simple model calculation and low material cost. Low, time and cost-effective

Pending Publication Date: 2022-04-05
鼎康(武汉)生物医药有限公司
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Problems solved by technology

[0005] The technical problem to be solved by the present invention is that the determination method of the dynamic capacity of the affinity chromatography filler in the prior art has the problems of long process time, large material consumption, long time consumption and high cost, or uses a relatively complicated calculation formula To calculate the dynamic load of affinity chromatography packing material, the processing period is long and the efficiency is low, thus providing a simple, clear, efficient, time-sensitive, and highly accurate method for the dynamic loading capacity of affinity chromatography packing material

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  • Determination method for dynamic loading capacity of affinity chromatography filler
  • Determination method for dynamic loading capacity of affinity chromatography filler
  • Determination method for dynamic loading capacity of affinity chromatography filler

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Embodiment 1

[0035] The method for determining the dynamic capacity of the affinity chromatography filler in this embodiment first utilizes the affinity chromatography filler to fit the sample concentration-ultraviolet absorbance value standard curve, specifically, including the following steps:

[0036] In step 1), the protein purified by affinity chromatography is diluted to the target protein concentration of 2.4 g / l in a normal fermentation broth with an affinity chromatography balance solution.

[0037] Step 2), prepare an affinity chromatography filler (Amsphere TM A3) Chromatographic column, use affinity chromatography equilibrium buffer (20mmol / L phosphate+100mmol / L NaCl, pH7.3±0.2, conductivity 13.0±2.0mS / cm) to replace the preservation in the affinity chromatography filler Solution (100mmol / L acetate+2% benzyl alcohol buffer, pH5.0±0.2, conductivity 4.5±1.0mS / cm) and equilibrated affinity chromatography filler.

[0038] Step 3), the sample prepared in step 1) is loaded into ste...

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Abstract

The invention discloses a method for measuring the dynamic loading capacity of an affinity chromatography filler, which comprises the following steps: 1) loading a sample onto a chromatographic column filled with the affinity chromatography filler to be screened, collecting flow-through liquid, and recording the loading volume; the sample loading volume is the total sample loading volume when the concentration of the penetrating protein sample corresponding to the ultraviolet absorption value reaches 5%-10% of the concentration of the target protein in the sample; the sample is a protein solution; and 2) calculating the dynamic loading capacity of the affinity chromatography filler according to the sample concentration-ultraviolet absorption value standard curve and the sample loading volume in the step 1). According to the property characteristics of affinity adsorption of the affinity filler and the antibody, an affinity adsorption kinetic model is constructed, the advantages of a traditional static binding capacity determination method and a current standard dynamic binding capacity determination method can be considered, and the method is short in time consumption, low in cost and high in accuracy.

Description

technical field [0001] The invention relates to a method for determining the dynamic capacity of an affinity chromatography filler. Background technique [0002] Screening of affinity chromatography media based on dynamic capacity is an important process development step in affinity chromatography. The traditional affinity chromatography filler screening is to determine the dynamic binding capacity, which is completely in accordance with the standard dynamic binding capacity determination method. Although the accuracy is high, it needs to consider factors such as different retention times and loading concentrations. The disadvantage is that the process time is long, the material consumption is large, the time is long, and the cost is high. [0003] The screening of another affinity filler is carried out by high-throughput technology, although it can save a lot of process development time and experimental samples, and save process development costs. However, the loading cap...

Claims

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Application Information

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IPC IPC(8): G01N30/74B01J20/281
Inventor 刘莹曾建成于淼郑子荣
Owner 鼎康(武汉)生物医药有限公司
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