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Method for purifying follicle stimulating hormone

A follicle-stimulating hormone and purity technology, which is applied to the preparation methods of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems of decreased ability, difficult control, complicated impurities, etc., and achieve the effect of improving efficiency and improving yield.

Pending Publication Date: 2022-08-05
LG CHEM LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the use of dye affinity chromatography has the advantage of being excellent in recovering the target protein by removing the medium components in the medium, it also has the disadvantage of a reduced ability to remove impurities originating from the host cell
These impurities are so diverse and complex that they cannot be identified as a
Therefore, there is a high risk of changes in the quality of the target protein and these changes are difficult to control
In addition, in order to further remove a large amount of impurities, complex purification steps are required after the capture step, resulting in a decrease in yield

Method used

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  • Method for purifying follicle stimulating hormone
  • Method for purifying follicle stimulating hormone
  • Method for purifying follicle stimulating hormone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1: Purification process

Embodiment 1-1

[0084] Example 1-1: Sequence of purification process

[0085] [Table 1]

[0086]

[0087]

[0088] As described above, in order to optimize the sequence of purification steps and processes for FSH, the FSH protein was purified by 15 different purification methods. Purification proceeds to Step 1, Step 2, or Step 3, depending on the experiment, and the purified solution from each step is collected.

[0089] Since the purified fluid in each step is used as the loading sample for the next step, pretreatment can be performed as needed (eg dilution or buffer exchange for AEX processes, salt addition for HIC processes, etc.). All basic process conditions for IAC, HIC and AEX purification were used in the same way.

Embodiment 1-2

[0090] Examples 1-2: Purification conditions for each step

[0091] The buffers and specific process conditions used in each purification process are summarized in Tables 2 to 4 below.

[0092] [Table 2] IAC process

[0093] step buffer used Injection volume (CV) balance 20mM Tris (pH 7.6) 7 inject Culture broth or purified solution from the previous step - balance 20mM Tris (pH 7.6) 5 cleaning 1 5mM Tris (pH 7.6) 7 cleaning 2 20mM Tris (pH 7.6) / 1M NaCl 5 cleaning 3 20mM Tris (pH 7.6) / 0.2M MgCl 2

7 elution 20mM Tris (pH 7.6) / 2M MgCl 2

7 CIP 0.5M acetic acid 5

[0094] [Table 3] HIC process

[0095] step buffer used Injection volume (CV) balance 20mM Tris (pH 7.6) / 2M NaCl 5 inject Culture broth or purified solution from the previous step - balance 20mM Tris (pH 7.6) / 2M NaCl 3 elution 20mM Tris (pH 7.6) 10 regeneration WFI 5 CIP ...

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Abstract

The present invention relates to a method for purifying follicle stimulating hormone in high yield and high purity, said method comprising an immunoaffinity chromatography (IAC) step.

Description

technical field [0001] The present invention relates to a method for purifying follicle-stimulating hormone with high yield and high purity. Background technique [0002] A variety of different protein therapeutics that can be used as various therapeutics have been used not only to study the production of recombinant proteins, but also to their clinical application and commercialization. [0003] One of these recombinant proteins, follicle-stimulating hormone (FSH), is produced by gonadotropins in the anterior pituitary gland, released into the bloodstream, and works with luteinizing hormone (LH) to control the Hormones for oocyte maturation and spermatogenesis in males. Human FSH is used to treat anovulatory women, to stimulate the development of multiple follicles (superovulation), and in preparation for assisted conception (eg, IVF, ICSI, GIFT, and CIFT). In addition, human FSH is used to stimulate follicular maturation in women with low or no FSH production and spermat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/59C07K1/18C07K1/22
CPCC07K1/22C07K14/59C07K1/18
Inventor 地晃宇吕琇宾薛三淑朴津亨方捐精
Owner LG CHEM LTD
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