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Method used for perfusion culture and purification through modification of chromatograph

A technology of perfusion culture and chromatography, which is applied in the field of separation and purification, can solve the problems that the harvest liquid cannot be processed continuously, and achieve the effect of facilitating large-scale industrial production, easy modification, and simple process

Inactive Publication Date: 2015-12-02
南月英
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a method for refitting a chromatograph for perfusion culture and purification, which solves the problem that the chromatograph cannot continuously process the harvest liquid, reduces downstream fixed costs and labor costs, and has two streams A function to run multiple columns in parallel

Method used

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  • Method used for perfusion culture and purification through modification of chromatograph
  • Method used for perfusion culture and purification through modification of chromatograph
  • Method used for perfusion culture and purification through modification of chromatograph

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Such as figure 1 , Figure 9d and Figure 9e As shown, a method for refitting a chromatograph for perfusion culture purification, wherein the method is: use two valves on the chromatograph, and when the two valves are seven-way valves 1 and 2, the Ports ⑤ and ⑦ of the first seven-way valve 1 are respectively connected to ports ③ and ① of the second seven-way valve 2, and port ⑤ of the second seven-way valve 2 is sealed , No. ⑦ port of the second seven-way valve 2 is the product outlet, which is connected to the detector of the system instrument, and No. ④ port of the two seven-way valves 1 and 2 are waste liquid ports. The two columns 3 and 4 are respectively installed between ports ② and ⑥ of the two seven-way valves. The system pump solution enters from port ① of the first seven-way valve 1, and the sample pump solution enters from port ③ of the first valve 1. The number port enters, and by switching the valve positions of the two seven-way valves 1 and 2, two flow...

Embodiment 2

[0043] Such as image 3 , Figure 9d , Figure 9e and Figure 9f As shown, a method for refitting a chromatograph for perfusion culture purification, wherein the method is: use a seven-way valve 1 and three eight-way valves 6, 7, 8 on the chromatograph instead of two valves , connect the port ③ and port ⑤ of the seven-way valve 1 with a tee and connect them to the upper end of the first column 3, and connect the lower end of the first column to the first eight-way valve 6, and connect the port of the seven-way valve 1 The ① port is connected to the upper end of the second column 4, the lower end of the second column 4 is connected to the second eight-way valve 7, and the ④ port of the seven-way valve 1 is connected to the ⑥ port with a tee and connected to the No. The upper ends of the three pillars 5 and the lower end of the third pillar 5 are connected to the third eight-way valve 8. The outlets ② of the three eight-way valves 6, 7, and 8 are waste liquid ports. The three...

Embodiment 3

[0052] Such as Figure 4As shown, a method for refitting a chromatograph for perfusion culture purification, wherein the method is: use five eight-way valves 6, 7, 8, 9, 10 on the chromatograph to replace two valves, and the column Two eight-way valves 9 and 10 are used at the upper end, and ports ①, ② and ③ of the fourth eight-way valve 9 are respectively connected to ports ③, ① and ② of the fifth eight-way valve 10, and are respectively connected through the three-way Connected to the upper ends of the first column 3, the second column 4 and the third column 5, and the lower ends of the first column 3, the second column 4 and the third column 5 are respectively connected to the first eight-way valve 6 , the second eight-way valve 7 and the third eight-way valve 8, the ports ③ and ④ of the first eight-way valve 6 are respectively connected to the upper end of the third column 5 and the second column 4 through a three-way , the port ③ and port ④ of the second eight-way valve ...

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Abstract

The invention relates to a method used for perfusion culture and purification through modification of a chromatograph. The method comprises the following steps: (1) an instrument which is provided with two parallel flow paths and operates two pillars simultaneously is modified on the basis of the chromatograph, one flow path and one pillar are used for feeding samples, and the other flow path and the other pillar are used for carrying out simultaneous balance elution; (2) a pillar is additionally arranged on the basis of step (1), the two flow paths are parallel, and every two pillars are parallel; (3) an instrument which is provided with two parallel flow paths and operates three pillars simultaneously is modified on the basis of the chromatograph, one flow path and one pillar are used for feeding the samples, another flow path and pillar are used for carrying out simultaneous balance elution, and the third pillar is used for receiving liquid flowing from another two pillars; and (4) the modification scheme of two parallel flow paths with amplification and universality is provided. According to the method, the modification is easy, the operation is convenient, the process is simple, the downstream process of perfusion culture is continuously implemented, the automatic operation is realized, the parallel amplification from trial production to normal production is realized, and the large-scale industrialized production is convenient.

Description

technical field [0001] The invention belongs to the field of separation and purification and the technical field of perfusion culture, and in particular relates to a method for refitting a chromatograph for perfusion culture and purification. Background technique [0002] Traditional fed-batch cell culture usually uses thousands to tens of thousands of liters of reaction tanks, the culture time is 7-21 days, and the product yield of different antibodies ranges from 2-6g / L. The perfusion culture technology can maintain a longer cell culture time by continuously adding fresh medium to the reactor and removing the exhausted medium at the same time, up to several months. [0003] The advantage of the perfusion culture process is that higher cell density can be obtained, and more product yields can be obtained at the reactor level of the same scale. In general, the operating concentration of perfusion culture is 10-30 times that of fed-batch culture. For example, using a 50L bio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/10C12N5/00
Inventor 南月英
Owner 南月英