CN110938135A - Solid-liquid separation method for suspension in pregnant mare serum gonadotropin crude product extraction process - Google Patents
Solid-liquid separation method for suspension in pregnant mare serum gonadotropin crude product extraction process Download PDFInfo
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/59—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g.hCG [human chorionic gonadotropin]; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
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Abstract
The invention relates to a solid-liquid separation method of suspension liquid in the extraction process of pregnant mare serum gonadotropin crude products. The method comprises the following steps: s1, taking pregnant mare serum, adding metaphosphoric acid solution 1 to adjust pH to 3.0-3.8 to obtain metaphosphoric acid/pregnant mare serum suspension; and S2, pumping the metaphosphoric acid/pregnant mare serum suspension into a plate-and-frame filter press, wherein the plate-and-frame filter press is filled with a pretreated filtering paperboard, and carrying out solid-liquid separation to obtain a protein clear solution. The pretreatment of the filter paper sheet in step S2 includes the steps of: soaking the filtering paper board in 0.05-0.2% (m/v) metaphosphoric acid solution 2 for 50-100min, draining, and placing into a plate-frame filter press. The invention adopts a plate-and-frame filter press to replace the traditional decanter centrifuge, and solves the bottleneck of carrying out solid-liquid separation on metaphosphoric acid/pregnant mare serum suspension on a large scale in the PMSG production process by pregnant mare serum. The method has the advantages of low labor intensity, high working efficiency, low energy consumption, high PMSG yield, high activity and the like.
Description
Technical Field
The invention belongs to the technical field of pregnant mare serum gonadotropin crude product production, and particularly relates to a solid-liquid separation method for suspension in a pregnant mare serum gonadotropin crude product extraction process.
Background
Pregnant mare serum gonadotropin (PMSG for short) is glycoprotein gonadotropin secreted by chorioallantoic cells of pregnant mare. PMSG has double activities similar to Follicle Stimulating Hormone (FSH) and Luteinizing Hormone (LH), but mainly takes the effect of FSH as the main part and has obvious follicle development promoting effect; meanwhile, the compound has certain functions of ovulation promotion and corpus luteum formation, and has the functions of promoting the development of sperm tubes and sexual cell differentiation for male animals. In practice, the PMSG can regulate and control the estrus of the female animals at the same time, realize controllable synchronous batch production, shorten the mating time, reduce the infertility probability and improve the propagation efficiency. At present, PMSG has been widely used for inducing estrus, synchronous estrus, superovulation or increasing ovulation rate of animals; meanwhile, the medicine also has good curative effect on ovarian hypoplasia or male spermatogenic decline and the like.
The raw material for producing PMSG is mare serum collected from 40-120 days of pregnancy, and the classical extraction method is as follows: first with metaphosphoric acid (HPO)3) Performing metaphosphate-protein reaction with serum to precipitate impure protein in the serum, and performing solid-liquid separation to obtain clear liquid; then, ethanol and clear liquid are used for carrying out 'ethanol-protein' classification to precipitate partial impure protein in the serum, then, the clear liquid is obtained by carrying out solid-liquid separation, and finally, the protein is precipitated by the ethanol to obtain a PMSG crude product.
In the small-scale PMSG production process, the solid-liquid separation method adopts a sedimentation type centrifugal separation method. Because the solid content of the suspension of the metaphosphoric acid-protein reaction is up to more than 9 percent, in practice, if a tubular centrifuge is used for separation, the rotary drum has small volume and limited solid content, and for materials with high solid content, frequent unloading and cleaning are needed in the production operation process, so the working efficiency is low; in addition, due to the particularity of particles in the suspension formed by the plasma protein, the disk centrifuge cannot solve the solid-liquid separation problem of the suspension formed by the metaphosphate-protein reaction, and the obtained clear liquid is still very turbid. With the development of intensive and large-scale high-efficiency animal husbandry, the demand of the market for PMSG is greatly increased, the daily processing capacity of pregnant mare plasma needs to be increased from 100kg to 2000kg, the traditional sedimentation type centrifugation method cannot meet the solid-liquid separation in plasma extraction, and the search for a large-scale solid-liquid separation method and equipment becomes necessary.
Disclosure of Invention
Aiming at the problems in the preparation process of PMSG in the prior art, the invention provides a method for carrying out large-scale solid-liquid separation of feed liquid in suspension by using a plate-and-frame filter press instead of a sedimentation centrifuge, which solves the bottleneck of solid-liquid separation in the process of producing PMSG from pregnant mare plasma, greatly improves the working efficiency, reduces the energy consumption, saves the process time, and plays a positive role in maintaining the bioactivity of PMSG.
The above object of the present invention is achieved by the following technical solutions:
a solid-liquid separation method of suspension liquid in the pregnant mare serum gonadotropin crude product extraction process comprises the following steps:
s1, taking pregnant mare serum, adding metaphosphoric acid solution 1 to adjust pH to 3.0-3.8 to obtain metaphosphoric acid/protein suspension;
s2, pumping the metaphosphoric acid/protein suspension into a plate-and-frame filter press, wherein the plate-and-frame filter press is filled with a pretreated filter paper board, and carrying out solid-liquid separation to obtain a protein clear liquid A;
preferably, the pretreatment of the filter paper sheet in step S2 includes the steps of: soaking the filter paper board in the metaphosphoric acid solution 2 for 50-100min, draining, and putting into a plate-frame filter press.
Preferably, the concentration of the metaphosphoric acid solution 2 is 0.05 to 0.2% (m/v) by mass.
Preferably, the concentration of the metaphosphoric acid solution 1 in the step S1 is 2 to 8% (m/v) by mass.
Preferably, metaphosphoric acid solution 1 is added to adjust the pH to 3.5.
Preferably, the plate and frame filter press is used for circularly cleaning the pretreated filter paper board with metaphosphoric acid solution 3 for 20-40min after being filled with the pretreated filter paper board.
Preferably, the concentration of the metaphosphoric acid solution 3 is 0.03 to 0.06% (m/v) by mass.
Preferably, the filtering pressure of the plate-and-frame filter press is 0.05-0.12 MPa.
Preferably, the pore size of the filter paper board is 1-20 μm.
Compared with the prior art, the invention has the beneficial effects that:
the invention adopts the plate-and-frame filter press to replace the traditional decanter centrifuge, solves the bottleneck of carrying out solid-liquid separation on metaphosphoric acid/protein suspension on a large scale in the PMSG (permanent magnet synchronous generator) production process of pregnant mare serum, and has the advantages of low labor intensity, high working efficiency, low energy consumption and the like compared with a centrifugal method. And the filter material is pretreated to eliminate the charges carried in the filter paper plate, thereby reducing the adsorption of target Protein (PMSG) and effectively improving the bioactivity and yield of PMSG crude products.
Drawings
Figure 1 is a schematic diagram of a plate and frame filter press filtration.
Wherein, 1, a metaphosphoric acid/protein suspension liquid storage tank, 2, a pump, 3, a plate-and-frame filter press, 4, a filter paper board, 5 and a protein clear liquid A liquid storage tank.
Detailed Description
In the following, some embodiments are described for the method for solid-liquid separation of a suspension during extraction of crude pregnant mare serum gonadotropins according to the present invention, however, these embodiments are exemplary and the present disclosure is not limited thereto. And the drawings used herein are for the purpose of illustrating the disclosure better and are not intended to limit the scope of the invention.
In some embodiments of the present invention, the solid-liquid separation method of the suspension during the extraction of crude pregnant mare serum gonadotropins comprises the following steps:
s1, taking pregnant mare serum, adding metaphosphoric acid solution 1 to adjust pH to 3.0-3.8 to obtain metaphosphoric acid/protein suspension;
and S2, pumping the metaphosphoric acid/protein suspension into a plate-and-frame filter press, wherein the plate-and-frame filter press is filled with a pretreated filtering paperboard, and carrying out solid-liquid separation to obtain a protein clear solution.
Pregnant mare serum contains many proteins and is in a wide variety, and PMSG has low concentration in pregnant mare plasma, so that a large amount of impure proteins need to be removed firstly, and PMSG is concentrated and enriched. The metaphosphoric acid solution contains a large amount of polymers with high charges and large molecular weight, the charges on the surfaces of protein colloidal particles can be effectively neutralized, hydration membranes on the surfaces of the protein colloidal particles are damaged, the protein colloidal particles are coagulated and precipitated, the pH value is adjusted to be between 3.0 and 3.8, most of hetero proteins in plasma of pregnant mares are precipitated, the solid content of a suspension after metaphosphoric acid-protein reaction can be up to more than 9 percent, and if the large-scale production of PMSG is involved, the working efficiency of treating the suspension by using a decanter centrifuge is greatly reduced, and the cost is greatly increased. The invention adopts the plate-and-frame filter press to process the suspension after the metaphosphoric acid-protein reaction, and utilizes the characteristics of large filtering area, good permeability of the filter material and large volume and solid content of the filter material of the plate-and-frame filter press to replace the common sedimentation centrifuge to carry out solid-liquid separation of large-scale feed liquid. And the filter paper board in the plate-and-frame filter press needs to be treated in advance before use: the commercially available filter paper board is soaked in the metaphosphoric acid solution 2 for 50-100 min. The filter paper board can eliminate the charges carried in the filter paper board after being soaked in metaphosphoric acid solution, thereby reducing the adsorption of target Protein (PMSG), and the filter paper board treated by metaphosphoric acid can also keep the bioactivity of PMSG. If a commercially available untreated filter paper board is used directly, both the yield and the activity of the final PMSG crude product are significantly reduced. The mass concentration of the metaphosphoric acid solution 2 for soaking the filter paper board is not too low or too high, the concentration is too low, the effect of treating the filter paper board is not obvious, if the concentration is too high, the pH value of the filter paper board is obviously reduced, the fiber structure of the filter paper board is damaged, and the mass concentration of the metaphosphoric acid solution 2 is selected to be 0.05-0.2% (m/v).
The invention adopts a plate-and-frame filter press containing special pretreatment filter paper boards to treat suspension after metaphosphoric acid-protein reaction, and overcomes the following defects of a centrifugal separation method: firstly, the shearing force influences the protein, and the shearing force generated by high-speed centrifugation can cause the change of the three-dimensional structure of the protein; secondly, the influence of heat production on the protein, the machine can generate heat in the high-speed centrifugation process, so that the protein is heated and denatured; in the actual production process, in order to reduce heat generated by high-speed centrifugation, the centrifuge needs to be matched with a high-performance compressor unit to provide a cold environment so as to reduce heat generated by high-speed rotation of a rotor in a cavity, the energy consumption generated by the compressor is added with the high power consumption of the centrifuge, and the energy consumption generated in the centrifugation process is very large; and thirdly, in the centrifugal operation, because the capacity of a single centrifugal machine is limited, a plurality of centrifugal machines are required to be started simultaneously, the labor intensity of production personnel is high, the safety risk is high, and in the operation process, the dripping, leaking and sprinkling of materials cannot be avoided. Therefore, the invention adopts the plate-and-frame filter press to process the suspension, solves the bottleneck of solid-liquid separation in the PMSG production process from pregnant mare plasma, greatly improves the working efficiency, reduces the energy consumption, saves the process time, plays an active role in maintaining the bioactivity of PMSG, and improves the bioactivity and the yield of PMSG crude products to a certain extent.
The filtering paper board is a deep filtering medium compounded by natural plant fibers, high-efficiency filter aid and cationic resin, and liquid impurities are removed by two mechanisms of mechanical interception and electrostatic adsorption. The present invention prefers filter paper sheets with a pore size of 1-20 μm. The paper board can be selected from HS series paper boards of Beijing Forster Ruite filtration technology limited or other commercially available filtration paper boards, and one or more of HS600, HS800, HS1000, HS1600, HS2000, HS4000, HS6000, HS7000, HS8000, HS9000 series paper boards or similar varieties are selected.
In other embodiments of the present invention, the concentration of the metaphosphoric acid solution 1 in step S1 is 2 to 8% (m/v) by mass.
In other embodiments of the invention, metaphosphoric acid solution 1 is added to adjust the pH to 3.5.
In other embodiments of the present invention, the plate and frame filter press operation comprises the steps of:
loading the pretreated filter paper board into a plate-and-frame filter press, and ensuring the sealing of a sealing ring in the loading process; maintaining the pressure of the plate-and-frame filter press at 1.5-2.0 MPa; starting the equipment, circularly cleaning the pretreated filter paperboard by using a metaphosphoric acid solution 3 for 20-40min, wherein the mass concentration of the metaphosphoric acid solution 3 is 0.03-0.06% (m/v), and carefully checking whether a pipeline has a liquid leakage phenomenon when the equipment runs; opening an inlet end valve, starting a plate-and-frame filter press, pumping metaphosphoric acid/protein suspension, and collecting the filtered protein clear liquid A into a storage tank, wherein the filtering pressure is not more than 0.12MPa, preferably 0.05-0.12MPa, in the running process of equipment; and when the metaphosphoric acid/protein suspension is filtered, stopping the equipment, loosening the plate frame, and then removing the slag and cleaning by the filter press.
The schematic diagram of filtration by using a plate-and-frame filter press is shown in figure 1, metaphosphoric acid/protein suspension in a liquid storage tank (1) is pumped into a plate-and-frame filter press (3) by using a pump (2), and liquid is collected into a protein clear liquid A liquid storage tank (5) through a filter paper board (4).
The whole extraction process of pregnant mare serum gonadotropin crude products comprises the following steps:
s1, taking pregnant mare serum, adding metaphosphoric acid solution 1 to adjust pH to 3.0-3.8 to obtain metaphosphoric acid/protein suspension;
and S2, pumping the metaphosphoric acid/protein suspension into a plate-and-frame filter press, wherein the plate-and-frame filter press is filled with a pretreated filtering paperboard, and carrying out solid-liquid separation to obtain a protein clear liquid A. The filter board pretreatment comprises the following steps: soaking the filter paper board in the metaphosphoric acid solution 2 for 50-100 min.
S3, concentrating the protein clear liquid A, adding ethanol to enable the ethanol concentration of the solution system to be 45-60% (v/v), and performing solid-liquid separation to obtain a protein clear liquid B;
s4, adding ethanol into the protein clear liquid B to enable the ethanol concentration of the solution system to be 75-85% (v/v), standing for layering, siphoning to remove the supernatant, collecting the lower-layer precipitate, and drying in vacuum to obtain the pregnant mare serum gonadotropin crude product.
As for the solid-liquid separation technique in step S3, there is no limitation, and a decanter centrifuge or a plate and frame filter press may be used. And the standing and layering time in the step S4 is 10-24h, so that the upper layer and the lower layer are sufficiently separated, and the subsequent siphoning is facilitated.
Hereinafter, the technical solution of the present invention will be further described and illustrated by specific examples. Unless otherwise specified, the raw materials used in the following specific examples of the present invention are those commonly used in the art, and the methods used in the examples are those conventional in the art.
The centrifuge used in the following specific examples was a DL-8M ultra-large capacity refrigerated centrifuge (shanghai luxiang instrument centrifuge, ltd); the plate-and-frame filter press is a Niro 600 paperboard filter press of Beijing Forster Ruite filtration technology GmbH; metaphosphoric acid is chemically pure and purchased from chemical reagents of national medicine group, Inc.; pregnant mare serum is obtained by processing plasma collected from pregnant mare for 40-120 days.
Example 1
The process for extracting the PMSG crude product comprises the following steps:
120Kg of pregnant mare serum is taken and 5% (m/v) HPO is dripped3Adjusting pH to 3.5, suspending for 5min, pouring the obtained metaphosphoric acid/protein suspension into a centrifuge, centrifuging at 3000rpm for 30min, and separating to obtain protein clear liquid A; concentrating the protein clear liquid A to 30Kg with an ultrafiltration machine, then dropwise adding absolute ethyl alcohol to make the ethanol concentration of the system reach 50%, and carrying out centrifugal separation on the 50% ethanol-protein suspension at the centrifugal speed of 3000rpm for 10min to obtain protein clear liquid B55.3Kg; and (3) dropwise adding absolute ethyl alcohol into the protein clear liquid B to enable the concentration of the ethyl alcohol in the system to reach 80%, standing for 12h for layering, siphoning to remove the supernatant, collecting the lower-layer precipitate, and drying in vacuum to obtain 96.8g of PMSG crude product.
Example 2
The process for extracting the PMSG crude product comprises the following steps:
120Kg of pregnant mare serum is taken, 5% (m/v) metaphosphoric acid solution is dripped to adjust the pH value to 3.5, and metaphosphoric acid/protein suspension is obtained after 5min of suspension.
Selecting an HS7000 filter paper board, and soaking the filter paper board for 60min by using 0.1% (m/v) metaphosphoric acid solution; loading the pretreated filter paper board into a plate-and-frame filter press; performing pressure maintaining on a plate-and-frame filter press, wherein the pressure maintaining pressure is 2.0 MPa; starting the device, and circularly cleaning the pretreated filter paper board for 30min by using 0.05% (m/v) metaphosphoric acid solution; opening the inlet valve, starting the plate-and-frame filter press, pumping the metaphosphoric acid/protein suspension, setting the filtration pressure to be 0.10MPa in the running process of the equipment, and filtering to obtain protein clear liquid A.
Concentrating the protein clear liquid A to 30Kg with an ultrafilter, then dropwise adding anhydrous ethanol to make the ethanol concentration of the system reach 50%, and carrying out centrifugal separation on the 50% ethanol-protein suspension at the centrifugal speed of 3000rpm for 10min to obtain 56.4Kg of protein clear liquid B; and (3) dropwise adding absolute ethyl alcohol into the protein clear liquid B to enable the concentration of the ethyl alcohol in the system to reach 80%, standing for 12h for layering, siphoning to remove the supernatant, collecting the lower-layer precipitate, and drying in vacuum to obtain 92.6g of PMSG crude product.
Example 3
The process for extracting the PMSG crude product comprises the following steps:
300Kg of pregnant mare serum is taken, 4% (m/v) metaphosphoric acid solution is dripped to adjust the pH value to 3.5, and after suspension for 6min, metaphosphoric acid/protein suspension is obtained.
Selecting HS8000 filtering cardboard, soaking the filtering cardboard for 70min by 0.12% (m/v) metaphosphoric acid solution; loading the pretreated filter paper board into a plate-and-frame filter press; performing pressure maintaining on the plate-and-frame filter press, wherein the pressure maintaining pressure is 1.8 MPa; starting the device, and circularly cleaning the pretreated filter paper board for 30min by using 0.04% (m/v) metaphosphoric acid solution; opening the inlet valve, starting the plate-and-frame filter press, pumping the metaphosphoric acid/protein suspension, setting the filtration pressure to be 0.11MPa in the running process of the equipment, and filtering to obtain protein clear liquid A.
Concentrating the protein clear liquid A to 75Kg with an ultrafilter, then dropwise adding anhydrous ethanol to make the ethanol concentration of the system reach 50%, and carrying out centrifugal separation on the 50% ethanol-protein suspension at the centrifugal speed of 3000rpm for 10min to obtain the protein clear liquid B138.7Kg; and (3) dropwise adding absolute ethyl alcohol into the protein clear liquid B to enable the concentration of the ethyl alcohol in the system to reach 80%, standing for 12h for layering, siphoning to remove the supernatant, collecting the lower-layer precipitate, and drying in vacuum to obtain 223.4g of PMSG crude product.
Example 4
The process for extracting the PMSG crude product comprises the following steps:
900Kg of pregnant mare serum is taken, 6 percent (m/v) metaphosphoric acid solution is dripped to adjust the pH value to 3.5, and the metaphosphoric acid/protein suspension is obtained after suspension for 6 min.
Selecting an HS8000 filter paper board, and soaking the filter paper board for 80min by using 0.09% (m/v) metaphosphoric acid solution; loading the pretreated filter paper board into a plate-and-frame filter press; performing pressure maintaining on a plate-and-frame filter press, wherein the pressure maintaining pressure is 1.9 MPa; starting the device, and circularly cleaning the pretreated filter paper board for 30min by using 0.03% (m/v) metaphosphoric acid solution; and opening the inlet valve, starting the plate-and-frame filter press, pumping the metaphosphoric acid/protein suspension, setting the filtering pressure to be 0.09MPa in the running process of the equipment, and filtering to obtain protein clear liquid A.
Concentrating the protein clear liquid A to 225Kg by using an ultrafiltration machine, then dropwise adding absolute ethyl alcohol to ensure that the ethanol concentration of the system reaches 50%, and carrying out centrifugal separation on the 50% ethanol-protein suspension at the centrifugal speed of 3000rpm for 10min to obtain 420.8Kg of protein clear liquid B; and (3) dropwise adding absolute ethyl alcohol into the protein clear liquid B to enable the concentration of the ethyl alcohol in the system to reach 80%, standing for 12h for layering, siphoning to remove the supernatant, collecting the lower-layer precipitate, and drying in vacuum to obtain 633.6g of PMSG crude product.
Example 5
The process for extracting the PMSG crude product comprises the following steps:
and (3) taking 1500Kg of pregnant mare serum, dropwise adding 5% (m/v) metaphosphoric acid solution to adjust the pH to 3.5, and suspending for 6min to obtain metaphosphoric acid/protein suspension.
Selecting HS9000 filter paper board, and soaking the filter paper board in 0.1% (m/v) metaphosphoric acid solution for 60 min; loading the pretreated filter paper board into a plate-and-frame filter press; performing pressure maintaining on a plate-and-frame filter press, wherein the pressure maintaining pressure is 2.0 MPa; starting the device, and circularly cleaning the pretreated filter paper board for 30min by using 0.04% (m/v) metaphosphoric acid solution; opening the inlet valve, starting the plate-and-frame filter press, pumping the metaphosphoric acid/protein suspension, setting the filtration pressure to be 0.10MPa in the running process of the equipment, and filtering to obtain protein clear liquid A.
Concentrating the protein clear liquid A to 300Kg with an ultrafilter, then dropwise adding anhydrous ethanol to make the ethanol concentration of the system reach 50%, and carrying out centrifugal separation on the 50% ethanol-protein suspension at the centrifugal speed of 3000rpm for 10min to obtain 546.2Kg of protein clear liquid B; and (3) dropwise adding absolute ethyl alcohol into the protein clear liquid B to enable the concentration of the ethyl alcohol in the system to reach 80%, standing for 12h for layering, siphoning to remove the supernatant, collecting the lower-layer precipitate, and drying in vacuum to obtain 1014.5g of PMSG crude product.
Example 6
Example 6 differs from example 2 in that the HS7000 filter board of example 6 was not soaked with 0.1% (m/v) metaphosphoric acid solution and the filter board was not washed with 0.05% (m/v) metaphosphoric acid solution after the plate and frame filter press was opened. The other process parameters are the same as those in example 2, and 90.2g of PMSG crude product is obtained.
Example 7
Example 7 differs from example 2 in that the HS7000 filter board of example 7 was soaked for 30min with a 0.5% (m/v) metaphosphoric acid solution. Other process parameters are the same as those in example 2, and 95.7g of PMSG crude product is obtained.
Aiming at metaphosphoric acid/protein suspension produced by metaphosphoric acid-protein reaction, in the production process, each ton of serum is fed, if a plate-and-frame filter press is used, one plate-and-frame filter press with the filtering area of 30 square can finish filtering within about 80 minutes, which is equivalent to the working capacity of 50 DL-8M super-large-capacity refrigerated centrifuges within 80 minutes. The energy consumption of the present invention using a plate and frame filter press can be greatly reduced as compared to the energy consumption of table 1 below.
TABLE 1 energy consumption comparison of solid-liquid separation apparatus
| Plate frame filter press | Ultra-large capacity centrifuge | |
| Number of | 1 table | 50 stands |
| Electric power | 7.5 kilowatt/station | 7.5 kilowatt/station |
| Total power | 7.5 kilowatts | 375 kw |
The activity data for the crude PMSG of inventive examples 1-7 are shown in table 2 below. In table 2, the PMSG yield was calculated in the following manner:
yield% (% crude total activity/total plasma activity)% 100%
Table 2 bioactivity data for examples 1-7
As can be seen from the data in Table 1, the solid-liquid separation of the large-capacity suspension can be processed with high efficiency and low consumption in the examples 2-5 by adopting the plate-and-frame filter press for filtration, and the activity and yield of the obtained PMSG crude product are higher than those of the PMSG crude product obtained by the centrifuge for filtration. While the pretreatment of the filter paper board plays a great role in crude extraction of PMSG, in example 6, the filter paper board without pretreatment is adopted, and although the efficiency is higher and the consumption is lower compared with the centrifugal method, the filter paper board without metaphosphoric acid treatment has a certain adsorption effect on target protein PMSG, so that the yield and the activity of the final extracted crude product are reduced.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (9)
1. A solid-liquid separation method of suspension liquid in the pregnant mare serum gonadotropin crude product extraction process is characterized by comprising the following steps:
s1, taking pregnant mare serum, adding metaphosphoric acid solution 1 to adjust pH to 3.0-3.8 to obtain metaphosphoric acid/protein suspension;
and S2, pumping the metaphosphoric acid/protein suspension into a plate-and-frame filter press, wherein the plate-and-frame filter press is filled with a pretreated filtering paperboard, and carrying out solid-liquid separation to obtain a protein clear solution.
2. The method for solid-liquid separation of suspension liquid during extraction of crude pregnant mare serum gonadotropin as claimed in claim 1, wherein the pretreatment of the filter paper board in step S2 comprises the following steps: soaking the filter paper board in the metaphosphoric acid solution 2 for 50-100 min.
3. The method for separating solid from liquid in suspension liquid during the extraction process of pregnant mare serum gonadotropin crude product as claimed in claim 2, wherein the mass concentration of the metaphosphoric acid solution 2 is 0.05-0.2% (m/v).
4. The method for separating solid from liquid in suspension liquid during the extraction process of pregnant mare serum gonadotropin crude product as claimed in claim 1, wherein the mass concentration of the metaphosphoric acid solution 1 in step S1 is 2-8% (m/v).
5. The method of claim 1, wherein the pH of the suspension is adjusted to 3.5 by adding metaphosphoric acid solution 1.
6. The method for separating solid and liquid from suspension liquid in the process of extracting crude pregnant mare serum gonadotropin as claimed in claim 1, wherein the plate and frame filter press is used for circularly cleaning the pretreated filtering paper board with metaphosphoric acid solution 3 for 20-40min after being filled with the pretreated filtering paper board.
7. The method for separating solid from liquid in suspension liquid during the extraction process of pregnant mare serum gonadotropin crude product as claimed in claim 6, wherein the mass concentration of the metaphosphoric acid solution 3 is 0.03-0.06% (m/v).
8. The method for separating solid from liquid in the process of extracting crude pregnant mare serum gonadotropin as claimed in claim 1, wherein the filtering pressure of the plate and frame filter press is 0.05-0.12 MPa.
9. The method for solid-liquid separation of suspension during extraction of crude pregnant mare serum gonadotropin as claimed in claim 1, wherein the pore size of the filter paper board is 1-20 μm.
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| CN207294655U (en) * | 2017-08-04 | 2018-05-01 | 蒙马（天津）生物科技有限公司 | A kind of extraction system of pregnant mare serum gonadotrop(h)in (PMSG) |
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