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Simulated moving bed chromatographic focusing

A chromatographic column and injection tube technology, which is applied in the field of invention of different individual components in their respective regions, can solve problems such as low concentration of fractions and improve the problem, and achieve the effects of increasing product yield and purity, increasing yield, and reducing production consumption

Inactive Publication Date: 2008-12-24
ODEKO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This patent focuses on how to separate the three components in the mixture, and does not improve the low concentration of the distillate

Method used

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  • Simulated moving bed chromatographic focusing
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  • Simulated moving bed chromatographic focusing

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0061] Example 1: Batch gradient chromatography separation of porcine serum

[0062] A chromatographic column with an inner diameter of 4.6 mm and a length of 180 mm was filled with DEAE-Sepharose ion exchange resin (particle size 50 μm, from Pharmacia). The sample is 10mM KCl semi-permeable membrane filtered porcine serum. Add 1ml of sample to the chromatographic column, then elute the chromatographic column with 30ml linear gradient KCl solution (concentration from 0.01M to 0.4M), and collect 30 fractions. These fractions were analyzed by 10% non-modified polyacrylamide gel electrophoresis in the Laemmli buffer system. The greatest amount of albumin occurs in fractions 14-20. A weak band (named component B) co-eluted with albumin in fractions 17 and 18.

example 2

[0063] Example 2: Separation of porcine serum by simulated moving bed focusing chromatography (hereinafter referred to as SMBCF)

[0064] Such as Figure 5 The form shown constitutes the SMBCF system. Unit F includes 5 areas, and Unit S, Unit B, and Unit R each include one area. There is a chromatographic column (30 mm×4.6 mm, length×inner diameter) in each zone, filled with DEAE-Sepharose ion exchange resin (50 μm, from Pharmacia). system such as Figure 5 Operate as described. Perform a column reset every 1.5 minutes. Pig serum was treated as a sample in the same manner as in Example 1, and see Table 1 for the composition and injection flow rate of the eluate in all 8 zones. The system performs the column reset process 40 times. Each time the chromatographic column is reset, collect all the eluates from the S(1) and F(1) regions; collect 0 from each of the F(2), F(3), F(4), and F(5) regions. .025ml of eluate. Fractions collected at the 10th, 20th, 30th, and 40th colu...

example 3

[0067] Example 3: Separation of SMBCF from porcine serum with fluid harvesting by fluid displacement

[0068] Such as Figure 5 The composition of the SMBCF system is shown. Use the method of Example 2 to separate. However, the liquid collection in the F(3) area is used Figure 8The flow displacement shown was carried out. Liquid was collected every 5 column resets. 0.05ml of 0.17M NaCl solution was used as the replacement solution for each collection. The other liquids were received in the same manner as in Example 2. Fractions collected at the 10th, 20th, 30th, and 40th column resets were analyzed using 10% non-modified polyacrylamide gel electrophoresis under the Laemmli buffer system. Compared with Example 2, the band of component B in the fraction collected in the F(3) area is stronger and the purity is higher.

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Abstract

A continuous simulated moving bed focusing chromatography method disclosing the separation and concentration of multiple components from a complex compound. This method consists of several zones connected together in a certain order, and the eluent is eluted unidirectionally along these zones, and is serially diluted one zone after another to reduce its desorption strength; each zone is equipped with A detachable column that periodically resets upstream one zone. The sample containing multiple components is injected into the downstream zone and adsorbed in the chromatography column in this zone, and is brought upstream zone by zone with the reset of the chromatography column, and is selectively desorbed from different zones And be separated. A certain component desorbed from one zone is re-adsorbed in the chromatography column in the adjacent downstream zone, and is brought back to the previous zone as the chromatography column is reset. The sample is thus maintained between the preceding zone and its adjacent downstream zone, and is accumulated by successive loadings and repeated resetting of the column.

Description

technical field [0001] The invention relates to a method for continuous chromatography using a multi-zone simulated moving bed structure. Rather, it is an invention that focuses on separating and concentrating the different individual components of a mixture in their respective regions. Background technique [0002] Chromatographic separation can be divided into two ways: batch chromatography and continuous chromatography. Most routine chromatography is performed in batch chromatography. Typically, batch chromatography separates samples in batches. The separation process of each batch consists of the same steps of sample loading, elution and fraction collection, and chromatographic column regeneration. The advantage of batch chromatography is that it can be used to separate complex compounds. However, in order to ensure a good separation effect, the sample loading should be less than 1% of the total column volume. This is one of the defects of the batch chromatography m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D15/08
Inventor 马起凤
Owner ODEKO INC
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