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Process of purification of hCG and recombinant hCG purified by that method

A chorionic gonadotropin and hormone technology, which is used in the preparation methods of peptides, chemical instruments and methods, sexual diseases, etc.

Inactive Publication Date: 2009-04-29
MERCK SERONO SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Residual contaminants can be further removed using a size exclusion column

Method used

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  • Process of purification of hCG and recombinant hCG purified by that method

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Experimental program
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Embodiment 1

[0031] Reagent

[0032] Ammonia, analytical grade

[0033] Ammonium bicarbonate, analytical grade (British Pharmacopoeia)

[0034] Disodium hydrogen phosphate, analytical grade

[0035] absolute denatured ethanol

[0036] Phosphoric acid, analytical grade (European Pharmacopoeia)

[0037] Isopropanol, analytical grade (European Pharmacopoeia)

[0038] Sodium Chloride, Analytical Grade (European Pharmacopoeia)

[0039] Sodium dihydrogen phosphate, analytical grade

[0040] Sodium Hydroxide Granules, Analytical Grade (European Pharmacopoeia)

[0041] Trifluoroacetic acid (TFA), HPLC grade

[0042] Tris-(hydroxymethyl)aminomethane, analytical grade

[0043] Schematic diagram of the purification process

[0044] The harvest from the cell culture process was purified and concentrated by 5 consecutive chromatographic steps.

[0045] The following schematic flow chart (Table 1) is a preferred embodiment of the recombinant human chorionic gonadotropin (r-hCG) purification pr...

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Abstract

A process for the purification of recombinant human Chorionic Gonadotropin (hCG) from a sample of crude recombinant hCG in the supernatant of CHO cells comprises the combined use of ion-exchange chromatography and reverse phase HPLC. The ion-exchange chromatography is performed twice and the final use of a size exclusion chromatography allows the purification from any residual traces of contaminants. The specific bioactivity of the highly purified hCG obtained from the process is particularly high, amounting to about 25.000 IU / mg.

Description

technical field [0001] The invention relates to a method for purifying chorionic gonadotropin (CG), in particular to a method for purifying recombinant human chorionic gonadotropin from an original recombinant human chorionic gonadotropin sample. The method involves the use of ion exchange chromatography and reversed-phase high performance liquid chromatography. Background technique [0002] Chorionic gonadotropin (CG) is a hormone produced by the placenta and is usually extracted from the urine of pregnant women. [0003] The hormone is a heterodimer composed of α and β subunits bound non-covalently. [0004] It mainly affects the luteinizing hormone in the gonadotropins. [0005] In the treatment of anovulatory infertility due to the absence or low concentration of gonadotropins, women may be given chorionic gonadotropin after induction of follicle development stimulated by follicle-stimulating hormone or human menopausal gonadotropin ovulation. A single dose of 5,000 ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/36C07K14/59G01N30/26A61K38/24A61P15/08B01J20/281B01J20/283C12N15/09G01N30/34G01N30/46G01N30/88G01N30/96
CPCC07K14/59A61P15/00A61P15/08A61P5/00C07K1/36
Inventor G·帕拉迪斯M·罗西L·斯卡利亚
Owner MERCK SERONO SA
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