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Pregnant mare serum gonadotropin (PMSG) purifying method

A technology of gonadotropin and pregnant horse serum, which is applied in the field of bioengineering, can solve the problems of low yield and achieve the effect of improving the recovery rate

Active Publication Date: 2013-09-04
CHIFENG BOEN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the metaphosphoric acid separation-salting-out-ethanol precipitation-dialysis method is the most widely used method. It has simple technology, low equipment requirements, low production cost and is suitable for mass production. Low rate, often less than 30%

Method used

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  • Pregnant mare serum gonadotropin (PMSG) purifying method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: a kind of purification method of pregnant horse serum gonadotropin comprises the following steps:

[0031] (1) Metaphosphoric acid precipitation: Slowly add 0.40mol / L metaphosphoric acid HPO to the pregnant horse serum while stirring 3 , when the pH value of the solution is 3.1, stop adding acid and stirring, and collect the precipitate and supernatant by filtration;

[0032] (2) Ethanol cleaning: add dehydrated alcohol equal to the weight of the precipitate to the precipitate collected in step (1), stir and filter, collect the ethanol cleaning solution, and discard the precipitate;

[0033] (3) octanoic acid precipitation: in the supernatant collected in step (1), add dropwise the sodium hydroxide of 2mol / L to adjust the pH value to 4.4; Add caprylic acid, after precipitation occurs, centrifuge and filter to collect the supernatant, and discard the precipitation;

[0034] (4) Salt-out precipitation: Combine the ethanol cleaning solution collected in ste...

Embodiment 2

[0039] Embodiment 2: a kind of purification method of pregnant horse serum gonadotropin comprises the following steps:

[0040] (1) Precipitation of metaphosphoric acid: Slowly add 0.5mol / L metaphosphoric acid HPO to pregnant horse serum while stirring 3 , when the pH value of the solution is 3.2, stop adding acid and stirring, and collect the precipitate and supernatant by filtration;

[0041] (2) Ethanol cleaning: add dehydrated alcohol equal to the weight of the precipitate to the precipitate collected in step (1), stir and filter, collect the ethanol cleaning solution, and discard the precipitate;

[0042] (3) octanoic acid precipitation: in the supernatant collected in step (1), add dropwise 2mol / L sodium hydroxide to adjust the pH value to 4.5; Add caprylic acid, after precipitation occurs, centrifuge and filter to collect the supernatant, and discard the precipitation;

[0043](4) Salt-out precipitation: Combine the ethanol cleaning liquid collected in step (2) and th...

Embodiment 3

[0048] Embodiment 3: a kind of purification method of pregnant horse serum gonadotropin comprises the following steps:

[0049] (1) Metaphosphoric acid precipitation: Slowly add 0.45mol / L metaphosphoric acid HPO to the pregnant horse serum while stirring 3 , when the pH value of the solution is 3.3, stop adding acid and stirring, and collect the precipitate and supernatant by filtration;

[0050] (2) Ethanol cleaning: add dehydrated alcohol equal to the weight of the precipitate to the precipitate collected in step (1), stir and filter, collect the ethanol cleaning solution, and discard the precipitate;

[0051] (3) octanoic acid precipitation: in the supernatant collected in step (1), add dropwise 2mol / L sodium hydroxide to adjust the pH value to 4.6; Add caprylic acid, after precipitation occurs, centrifuge and filter to collect the supernatant, and discard the precipitation;

[0052] (4) Salt-out precipitation: Combine the ethanol cleaning solution collected in step (2) w...

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Abstract

The invention discloses a PMSG purifying method. The method comprises the steps of metaphosphoric acid precipitation, ethanol cleaning, caprylic acid precipitation, salting-out precipitation, dialysis desalting, low-concentration ethanol precipitation, high-concentration ethanol precipitation and dialysis cleaning. The addition of the caprylic acid precipitation step allows difficult-precipitation albumins and other macromolecular other proteins to precipitate, the salting-out precipitation step increases the ammonium sulfate adding concentration and avoids the albumin precipitation problem, and the increase of the ammonium sulfate addition concentration improves the precipitation proportions of PMSG active substances and also improves the precipitation proportions of follicle-stimulating hormone (FSH) active substances, so the PMSG purity is increased, and the PMSG recovery rate is also increased.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for purifying serum gonadotropin of pregnant horses. Background technique [0002] Pregnant horse serum gonadotropin, namely PMSG (Pregnant Mare Serum Gonadotropin). Also known as equine chorionic gonadotropin. It is a hormone found in the blood serum of pregnant mares. It has the dual functions of follicle-stimulating hormone (FSH) and luteinizing hormone (LH). It can prevent and treat infertility in livestock, induce superovulation, control estrus synchronization, and promote multiple births. [0003] A large number of non-gonadotropic macromolecular proteins in the serum of pregnant horses have antigenicity, and injection into non-equine livestock can easily cause anaphylactic shock and death in severe cases, especially if repeated application is more likely to occur. It is precisely because of allergic reactions, epidemiological and other reason...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/575C07K1/36C07K1/30C07K1/34
Inventor 刘永祥刘永存
Owner CHIFENG BOEN PHARMA
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