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Preparation method of high purity serum gonadotrophin

A production method and high-purity technology, applied in the field of biopharmaceuticals, to achieve the effects of high purification efficiency, less loss, and easy mastery of the process

Inactive Publication Date: 2007-02-28
NINGBO SANSHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there are research reports on the purification of PMSG by SE-Sephadx C-50, but SE-Sephadx C-50 itself has been eliminated

Method used

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  • Preparation method of high purity serum gonadotrophin

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0075] DEAE-Sephadex A-50 is an ion-exchange chromatography medium, first soaked in water for injection for 24 hours, then equilibrated to pH7.0 with initial buffer solution (0.01mol / L sodium acetate buffer solution of pH7.0), and loaded after equilibrium Column, the column is φ50mm×500mm, equilibrated with the initial buffer again.

[0076] The sample volume is about 2 million u of the crude product (potency is about 800 units / mg), and the crude product is dissolved in 100ml of 0.01mol / L sodium acetate buffer solution with a pH of 7.0, and the sample is slowly added dropwise after it is completely dissolved.

[0077] The amount of each gradient eluent is 3 times the volume of the column. Under the conditions of a flow rate of 10ml / min and a detection wavelength of 280nm, sequentially use the first gradient eluent (0.01mol / L sodium acetate buffer at pH 7.0) , the second gradient buffer (0.05mol / L sodium acetate buffer of pH7.0) and the third gradient buffer (0.2mol / L sodium ac...

preparation example 2

[0080] DEAE-Sephadex A-50 is an ion-exchange chromatography medium, which is first soaked in water for injection for 24 hours, then balanced to pH6.5 with an initial buffer solution (0.005mol / L sodium acetate buffer solution of pH6.5), and loaded Column, the column is φ50mm×500mm, equilibrated with the initial buffer solution again.

[0081] The sample volume is about 1.5 million u crude product (potency is about 650 units / mg), the crude product is dissolved in the initial buffer solution, and the sample is slowly added dropwise after complete dissolution.

[0082] The amount of each gradient eluent is 4 times the volume of the column. Under the conditions of a flow rate of 10ml / min and a detection wavelength of 280nm, the first gradient eluent (initial buffer solution) and the second gradient buffer (pH6. 5 0.07mol / L sodium acetate buffer) and the third gradient buffer (pH6.5 0.3mol / L sodium acetate buffer) elution process, detected by the nucleic acid protein detector, colle...

preparation example 3

[0085] DEAE-Sephadex A-50 is an ion-exchange chromatography medium, which is first soaked in water for injection for 24 hours, then balanced to pH8.5 with an initial buffer solution (0.015mol / L sodium acetate buffer solution of pH8.5), and loaded Column, the column is φ50mm×300mm, equilibrated with the initial buffer solution again.

[0086] The sample volume is about 2 million u crude product (potency is about 860 units / mg), the crude product is dissolved with the initial buffer solution, and the sample is slowly added dropwise after complete dissolution.

[0087] The amount of each gradient eluent is twice the volume of the column. Under the conditions of a flow rate of 10ml / min and a detection wavelength of 280nm, the first gradient eluent (initial buffer solution) and the second gradient buffer (pH8. 5 0.025mol / L sodium acetate buffer) and the third gradient buffer (0.15mol / L sodium acetate buffer of pH 8.5) during the elution process, after detection by a nucleic acid pro...

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Abstract

The invention discloses a preparing method of high-purity lobulantina, which is characterized by the following: adopting chromatography medium of DEAE-Sephadex A-50 as ionic exchange chromatography column, using PH as 6.5-8.5 and density is 0.005-0.07mol / L sodium acetate buffer solution to dissolve PMSG semifinished product and up-sample, using PH is 6.5-8.5 and density is 0.005-0.07mol / L sodium acetate buffer solution to break so as to remove foreign matter, using PH is 6.5-8.5 and density is 0.15-0.3mol / L sodium acetate buffer solution to elute and check at the some time, assembling eluent to get collecting solution, dialyzing and freeze-drying collecting solution to get elaboration product.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals in the field of medicine, in particular to a production method of high-purity gonadotrophin. Background technique [0002] PMSG is a glycoprotein gonadotropin secreted by the trophoblast cells of the placental uterus of pregnant horses. Since the discovery of PMSG in 1943, extensive technological research and application tests have been carried out. In the 1960s, D. Gospodarowicz and Papkoff in the United States established a relatively simple purification and separation technology, and at the same time carried out application tests for the treatment of infertility and infertility in livestock, and achieved good results. The Netherlands Intervet company started industrial production research in the mid-1960s, and it was officially applied to veterinary clinics in the 1970s. In the late 1970s, with the development of biotechnology, it was widely used in superovulation and estrus synchronization,...

Claims

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Application Information

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IPC IPC(8): C07K1/18C07K14/435
Inventor 俞通泰徐震宇翁士乔曾宏杰
Owner NINGBO SANSHENG PHARMA
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