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A kind of preparation technology of high-purity pregnant horse serum gonadotropin

A technology of gonadotropin and pregnant horse serum, which is applied in the preparation method of peptides, peptides, organic chemistry, etc., can solve the problems of low potency and poor quality, and achieve the effects of high potency, low risk of harm, and high purity

Active Publication Date: 2021-08-31
NINGBO SANSHENG BIOLOGICAL TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the defects of low titer and poor quality in the PMSG preparation process in the prior art, the present invention provides a PMSG preparation process to obtain high-purity PMSG with a titer ≥ 1500IU / mg

Method used

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  • A kind of preparation technology of high-purity pregnant horse serum gonadotropin
  • A kind of preparation technology of high-purity pregnant horse serum gonadotropin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The preparation process of pregnant horse serum gonadotropin of the present embodiment comprises the following steps:

[0026] (1) PMSG crude product preparation: add 1.5% metaphosphoric acid to adjust the pH of pregnant horse plasma to 3.8, stir for 10 minutes, centrifuge and filter, and the filtrate is concentrated to 1 / 2 volume, then add -20 ℃ absolute ethanol to make the ethanol concentration of the solution system 45% (v / v), stirred for 10 minutes, centrifuged, added -20 ℃ absolute ethanol to the filtrate to make the concentration of alcohols in the solution system 83% (v / v), stirred for 20 minutes, allowed to stand for 12 hours to separate layers , The supernatant was discarded by siphoning, and the lower precipitate was collected and vacuum-dried for 48 hours to obtain crude PMSG.

[0027](2) Preparation of PMSG intermediate product: add 10% (m / v) sodium 4-hydroxybenzoate solution to the crude product of PMSG, the mass ratio of the crude product of PMSG and sodiu...

Embodiment 2

[0030] The preparation process of pregnant horse serum gonadotropin of the present embodiment comprises the following steps:

[0031] (1) PMSG crude product preparation: add 1.5% metaphosphoric acid to adjust the pH of pregnant horse plasma to 3.8, stir for 10 minutes, centrifuge and filter, and the filtrate is concentrated to 1 / 2 volume, then add -20 ℃ absolute ethanol to make the ethanol concentration of the solution system 45% (v / v), stirred for 10 minutes, centrifuged, added -20 ℃ absolute ethanol to the filtrate to make the concentration of alcohols in the solution system 83% (v / v), stirred for 20 minutes, allowed to stand for 12 hours to separate layers , The supernatant was discarded by siphoning, and the lower precipitate was collected and vacuum-dried for 48 hours to obtain crude PMSG.

[0032] (2) Preparation of PMSG intermediate product: add 10% (m / v) sodium 4-hydroxybenzoate solution to the crude product of PMSG, the mass ratio of crude product of PMSG and sodium 4...

Embodiment 3

[0035] The preparation process of pregnant horse serum gonadotropin of the present embodiment comprises the following steps:

[0036] (1) PMSG crude product preparation: add 1% metaphosphoric acid to adjust the pH of pregnant horse plasma to 3.5, stir for 5 minutes, centrifuge and filter, and the filtrate is concentrated to 1 / 2 volume, then add -20 ℃ absolute ethanol to make the ethanol concentration of the solution system 40% (v / v), after stirring for 20 minutes, centrifuge, add -20 ℃ absolute ethanol to the filtrate to make the concentration of alcohols in the solution system 80% (v / v), after stirring for 20 minutes, let stand for 12 hours to separate layers , The supernatant was discarded by siphoning, and the lower precipitate was collected and vacuum-dried for 48 hours to obtain crude PMSG.

[0037] (2) Preparation of PMSG intermediate product: add 10% (m / v) sodium benzoate solution to PMSG crude product, the mass ratio of PMSG crude product and sodium benzoate is 1:0.1, ...

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PUM

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Abstract

A preparation process for high-purity pregnant horse serum gonadotropin, comprising the following steps: (1) adjusting the pH of pregnant horse plasma to 3.0-4.0, separating solid and liquid, concentrating the filtrate, and adding alcohols to make the alcohols in the solution system The concentration is 30-55% (v / v), and after solid-liquid separation, alcohols are added to the filtrate to make the concentration of alcohols in the solution system 70-90% (v / v), and the obtained precipitate is dried, which is PMSG Crude product; (2) Add toluene derivative or ethylbenzene derivative solution to the PMSG crude product, after solid-liquid separation, the precipitate is added with alcohol, and the obtained precipitate is dried, which is the PMSG intermediate product; (3) The PMSG intermediate product is added Ammonium salt and / or calcium salt and / or potassium salt solution and alcohol substances, after solid-liquid separation, the clear liquid is added to ammonium salt and / or calcium salt and / or potassium salt solution and alcohol substances for a second time, after solid-liquid separation , adding alcohols to the supernatant to obtain a precipitate and dry it to obtain a refined product of PMSG.

Description

technical field [0001] The invention belongs to the technical field of pregnant horse serum gonadotropin production, and in particular relates to a preparation process of high-purity pregnant horse serum gonadotropin. Background technique [0002] Pregnant horse serum gonadotropin (referred to as PMSG) is a glycoprotein gonadotropin secreted by the allantoic chorionic cells of pregnant mare. PMSG has dual activities similar to follicle-stimulating hormone (FSH) and luteinizing hormone (LH), but the effect of FSH is the main one, and it has obvious follicle-stimulating effect; meanwhile, it also has certain functions of ovulation-stimulating and corpus luteum formation, which is beneficial to male animals. It has the function of promoting the development of seminiferous tubules and the differentiation of sex cells. In practice, PMSG can regulate the simultaneous estrus of female animals, realize controllable and synchronous batch production, shorten mating time, reduce the p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/59C07K1/30
CPCC07K14/59
Inventor 俞通泰于钢沙乐王荣庆
Owner NINGBO SANSHENG BIOLOGICAL TECH CO LTD
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