CN110872345B - Preparation method of high-purity immunoglobulin G - Google Patents
Preparation method of high-purity immunoglobulin G Download PDFInfo
- Publication number
- CN110872345B CN110872345B CN201910770523.7A CN201910770523A CN110872345B CN 110872345 B CN110872345 B CN 110872345B CN 201910770523 A CN201910770523 A CN 201910770523A CN 110872345 B CN110872345 B CN 110872345B
- Authority
- CN
- China
- Prior art keywords
- immunoglobulin
- solution
- rutin
- virus
- uvc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/06—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from serum
- C07K16/065—Purification, fragmentation
Abstract
The invention provides a preparation method of high-purity immunoglobulin G, which separates the immunoglobulin G from animal serum by using a salting-out combined chromatography method and adopts an improved ultraviolet irradiation inactivation method for virus inactivation.
Description
Technical Field
The invention belongs to the field of protein and biological products, and particularly provides a preparation method of high-purity immunoglobulin G, which separates the immunoglobulin G from animal serum by using a salting-out combined chromatography method and adopts an improved ultraviolet irradiation inactivation method for virus inactivation.
Background
Immunoglobulin is the most abundant plasma protein except albumin in plasma, and accounts for about 12% -15% of the total amount of plasma protein. Mammalian immunoglobulins have high similarities in gene sequence, amino acid composition and protein structure, and their physiological functions and physiological actions are substantially identical: the basic unit of the immunoglobulin molecule is a symmetrical structure formed by four peptide chains, and comprises two identical heavy chains and two identical light chains, wherein the light chains are divided into two structural types of kappa and lambda; the main physiological functions and effects of the medicine are that the medicine can block or eliminate the pathogenic effect of various pathogens on the body by specifically binding corresponding antigens, activating complement and combining Fc receptors to generate antibody-dependent cell-mediated cytotoxicity and opsonophagocytosis, and the medicine can treat various diseases by antibody supplementation and immunoregulation.
The plasma immunoglobulin can be divided into five types, wherein the immunoglobulin G (IgG) accounts for about 70-80% of the total amount of the immunoglobulin, and is the most important immunoglobulin, the clinically used immunoglobulin product is mainly IgG, and the IgG infusion is a passive immunotherapy and is mainly applied to hypoimmunity, replacement therapy, prevention of certain viral and bacterial infections and the like.
The serum is light yellow liquid separated from whole blood collected from mammal blood vessel after natural coagulation, and is prepared into animal serum through further centrifugal separation; compared with plasma separated from anticoagulated whole blood, the serum has removed fibrinogen, various coagulation factors and other protein components, and the serum has fewer protein components, mainly including albumin, immunoglobulin and other trace proteins. Serum is a good raw material for preparing immune globulin
When serum immunoglobulin is purified, because raw materials such as plasma/serum contain a plurality of proteins, a single purification method often cannot meet the requirement, and a plurality of purification methods are combined usually, so that the purpose of purifying the immunoglobulin is achieved; at present, the purification method of medicinal immune globulin at home and abroad mainly takes the traditional low-temperature ethanol method as a main method, the method has high requirements on equipment, and the reaction steps need to be controlled at low temperature; the method has the advantages of multiple separation steps, long period, complex control of process conditions, huge energy consumption, and often generation of polymers in the purification process. In addition, viruses which can affect human bodies possibly existing in animal serum and the difficulty of animal screening make the virus inactivation process indispensable, a high-temperature heat treatment method, a low-pH treatment method and the like in common virus inactivation methods are not suitable for serum, currently used phenol methods and S/D methods have potential defects in the safety of residues and products, and photochemical treatment methods such as methylene blue have poor inactivation effect on non-enveloped viruses.
Therefore, there is still a great need in the art for a method for preparing immunoglobulin G having high purity, simple process, and good safety.
Disclosure of Invention
In order to solve the problems, the inventor adopts a salting-out precipitation separation method to replace a low-temperature ethanol separation method to carry out coarse purification on immunoglobulin G, and then carries out chromatographic separation on the primarily purified immunoglobulin G, thereby achieving the aim of quickly and simply preparing high-purity serum immunoglobulin G. In addition, when the inventor discovers that medicinal rutin with a specific proportion and a certain sodium ion concentration can be effectively inactivated by a UVC photochemical inactivation method under the condition of plasma virus inactivation (see the application on the same day in the application: rutin can be combined with virus nucleic acid and the virus nucleic acid can be effectively inactivated under the UVC illumination condition (500-5000 mu w/cm)2Irradiation for 120-1200 s) can make the virus nucleic acid produce gap and can not complete replication, so as to achieve the effect of inactivating virus; the method directly acts on virus nucleic acid, so the method has a good inactivation effect on both enveloped viruses and non-enveloped viruses, and further solves the problem that the methylene blue light chemical method can only inactivate the enveloped viruses and can not inactivate the non-enveloped viruses).
In one aspect, the present application provides a method for preparing high purity immunoglobulin G, which is characterized in that animal serum is used as a raw material, and the method comprises the steps of salting out and chromatographic separation.
Further, the animal serum is horse serum or dog serum.
Further, the preparation method of the high-purity immunoglobulin G comprises the following steps:
(1) diluting the raw material serum with normal saline until the protein concentration is less than or equal to 40 mg/mL;
(2) adding saturated ammonium sulfate solution at room temperature until the saturation reaches 30%, and centrifuging or filtering to remove impurity protein precipitate;
(3) collecting the supernatant or filtrate, adding saturated ammonium sulfate solution until saturation reaches 50%, centrifuging or filtering to collect immunoglobulin precipitate;
(4) dissolving immunoglobulin precipitate with 1-2 times of sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution, press-filtering with plate frame to remove insoluble impurities, further obtaining clear solution containing pure immunoglobulin G, and adjusting protein content to 5-10 mg/mL;
(5) inactivating viruses;
(6) ion exchange chromatography to obtain high purity immunoglobulin G solution.
Further, the adding amount of the physiological saline in the step (1) is 0.3-2 times of the volume of the serum; placing the diluted serum in a beaker in the step (2), starting stirring, controlling the stirring speed to be 20-60 rpm, slowly adding the prefabricated saturated ammonium sulfate solution according to the proportion until the saturation reaches 30%, controlling the reaction temperature to be less than 16 ℃, continuing stirring for 1h after the saturated ammonium sulfate solution is completely added, stopping the machine and standing for 1-3 h to ensure that the precipitation reaction is sufficient, carrying out centrifugal separation or filter pressing separation for solid-liquid separation after the precipitation reaction is completed, preferably carrying out filter pressing separation, collecting supernatant and discarding precipitated impure protein; placing the supernatant obtained by separation in the step (2) in a beaker, starting stirring, controlling the stirring speed to be 20-60 rpm, continuously and slowly adding the prefabricated saturated ammonium sulfate solution according to the proportion until the saturation reaches 50%, controlling the reaction temperature to be less than 16 ℃, continuously stirring for 1h after the saturated ammonium sulfate solution is completely added, stopping standing for 1-3 h, and fully performing precipitation reaction; after the precipitation reaction is finished, carrying out solid-liquid separation by centrifugal separation or filter-pressing separation, preferably filter-pressing separation, and collecting precipitates; in step (4), 1-5% of a filter aid selected from diatomaceous earth, activated carbon or perlite, preferably 3% diatomaceous earth, is used.
Further, in the step (6), an anionic chromatography medium selected from the group consisting of DEAE (N, N-diethylaminoethyl) as a ligand and Q (N, N-diethylamino-2-hydroxypropyl) as a ligand, preferably a chromatography medium having DEAE as a ligand, such as DEAE-sepharose Fast Flow, is used to directly load the solution after virus inactivation in the step (5).
Further, in the step (4), 40mM sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution of pH 6.9 was used for dissolution; in the step (5), 0.5g/L of medicinal rutin is added into the clear solution obtained in the step (4); obtaining the clear solution of the refined immunoglobulin G containing the rutin.
Further, the refined pure immunoglobulin G clear solution containing rutin flows through a spiral quartz glass tube, and the solution flowing through the glass tube is simultaneously subjected to UVC illumination with the intensity of 500-5000 mu w/cm2And (3) performing irradiation treatment for 120-1200 s.
The serum herein may be of various origins including, but not limited to, human, porcine, canine, bovine, equine, etc. origin.
The rutin, the phosphate, the ammonium sulfate and the chromatography medium in the application can adopt domestic or imported products which are compounded with relevant standards. Preferably a product which is standard for compound injection or standard for biological preparation.
The light irradiation treatment in the present application may be carried out by using various commercially available or commercially available light irradiation treatments which can provide a wavelength of 254nm and a wavelength of 500 to 5000. mu.w/cm2An illumination device for illumination intensity. For example, the inventors have made a spiral quartz glass tube with an inner diameter of 1 to 10mm, preferably 4 mm; the wall thickness of the spiral sample tube is 0.1 to 3mm, preferably 1mm (phi ═ 6. + -.1).
The salting-out chromatography method overcomes the defects of complex process steps, high operating condition requirement, severe operating environment, huge equipment investment, high energy consumption and the like of a low-temperature ethanol method, simultaneously avoids the generation of polymer protein in the ethanol precipitation process, and can greatly improve the quality of products; the improved UVC photochemical plasma inactivation method can effectively inactivate the enveloped viruses and the non-enveloped viruses, wherein the added low-concentration rutin and a small amount of sodium chloride are common components in the injection, the safety is relatively guaranteed, and various safety evaluations can be easily carried out.
Drawings
FIG. 1 is an ion exchange chromatogram of canine immunoglobulin G according to the present invention;
FIG. 2 is an ion exchange chromatogram of equine immunoglobulin G according to the present invention.
Detailed Description
The present invention will be described in detail with reference to specific examples, but the scope of the present invention is not limited to the following examples.
EXAMPLE 1 Primary reagents, raw materials and apparatus
Horse serum and dog serum were purchased from Beijing Tianqin I and Biotech, Inc.;
pseudorabies virus and encephalomyocarditis virus were purchased from the Chinese veterinary microbial strain preservation management center;
PK-15 and BHK-21 cells were purchased from China veterinary microbial culture collection management center;
the medicinal rutin is produced by Sichuan synergetic pharmaceutical products GmbH;
the ammonium sulfate is produced by Limited company of Xian Tianzheng pharmaceutic adjuvant;
phosphate is produced by national medicine group;
DEAE-sepharose Fast Flow for Amazonia (GE) production;
the rest reagents and instruments are all conventional domestic models.
Example 2 preparation of high purity Canine immunoglobulin G
1 after thawing the raw material dog serum, 200mL of the raw material dog serum was placed in a beaker, 100mL of physiological saline was added for dilution, and the pH of the diluted serum was adjusted to 7.0 with 1% NaOH.
2, placing the beaker on a magnetic stirrer, starting stirring, controlling the stirring speed at 50rpm, slowly adding 128mL of a prefabricated saturated ammonium sulfate (pharmaceutic adjuvant grade) solution, continuing stirring for 1h, determining the reaction temperature to be 14 ℃, stopping the machine and standing for 1 h; then solid-liquid separation is carried out by a centrifugal machine, supernatant is collected, precipitated protein impurities are discarded, centrifugal separation is adopted, and the supernatant is collected for standby.
3, placing the supernatant obtained by separation in the step (2) in a beaker, starting stirring, controlling the stirring speed at 50rpm, continuously and slowly adding 172mL of a prefabricated saturated ammonium sulfate (pharmaceutic adjuvant grade) solution according to the proportion, measuring the reaction temperature to be 15 ℃, continuously stirring for 1h after the addition is finished, stopping the machine, standing for 2h, and fully performing precipitation reaction; and after the precipitation reaction is finished, performing solid-liquid separation by using a centrifugal machine, and collecting the precipitate for later use.
4 dissolving the precipitate (immunoglobulin) collected in the step (3) with 200mL of a 40mM sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution having a pH of 6.9, filtering the solution through a microfiltration membrane (0.45 μm) to remove insoluble impurities, and further obtaining a clear solution containing purified canine immunoglobulin G, determining the protein content and adjusting the protein content to 6 mg/mL.
5, performing virus inactivation treatment, adding 0.5G/L medicinal rutin into the obtained clarified solution, enabling the clarified solution of the pure immune globulin G containing the rutin to flow through a helical quartz glass tube, and simultaneously subjecting the solution flowing through the glass tube to UVC illumination with the intensity of 500-5000 mu w/cm2And (3) performing irradiation treatment for 120-1200 s.
6 directly loading the reaction solution after virus inactivation treatment for chromatographic separation (see figure 1)
(1) 10mL of an anionic chromatography medium (DEAE-sepharose Fast Flow) was loaded into an XK16/20 column;
(2) equilibrating the column with 20mM PBS buffer (pH7.0);
(3) injecting a chromatography sample loading solution containing dog immunoglobulin G into a chromatography column through an AKTA pure chromatograph, controlling the flow rate to be 1mL/min, monitoring a protein ultraviolet absorption peak, and collecting penetration liquid;
(4) after the sample loading is finished, continuously performing flow washing by using 20mM PBS buffer solution (pH7.0), and stopping collecting until the ultraviolet absorption peak of the protein is reduced to a baseline;
(5) after the flow washing is finished, changing the flow washing into 20mM PBS +1M NaCl buffer solution (pH7.0), eluting the hybrid protein combined with the chromatographic medium, and regenerating the chromatographic column;
7, performing ultrafiltration on the collected chromatography penetration peak solution by using an ultrafiltration membrane with the molecular weight cutoff of 50kDa, and replacing the solution with a dog immunoglobulin storage buffer solution which is a glycine solution of 0.2 mol/L; the volume of the ultrafiltration liquid exchange solution is 10 solution volumes, and the solution is concentrated until the content of the canine immunoglobulin is about 25 mg/mL; filtering and sterilizing by using a 0.2 mu m sterilizing filter membrane to obtain the high-purity dog immunoglobulin G solution.
EXAMPLE 3 preparation of high purity equine immunoglobulin G
1 melting raw material horse serum, placing 200mL into a beaker, adding 100mL of normal saline for dilution, and adjusting the pH value of the diluted serum to 7.0 by using 1% NaOH.
2, placing the beaker on a magnetic stirrer, starting stirring, controlling the stirring speed at 50rpm, slowly adding 128mL of a prefabricated saturated ammonium sulfate (pharmaceutic adjuvant grade) solution, continuing stirring for 1h, determining the reaction temperature to be 14 ℃, stopping the machine and standing for 1 h; after the completion of the precipitation reaction, 13 g of a diatomaceous earth filter aid was added in an amount of 3%, solid-liquid separation was carried out using a microporous filter plate (model: SS335), and the supernatant was collected for use.
3, placing the supernatant obtained by separation in the step (2) in a beaker, starting stirring, controlling the stirring speed at 50rpm, continuously and slowly adding 172mL of a prefabricated saturated ammonium sulfate (pharmaceutic adjuvant grade) solution according to the proportion, measuring the reaction temperature to be 14 ℃, continuously stirring for 1h after the addition is finished, stopping the machine, standing for 2h, and fully performing precipitation reaction; after the completion of the precipitation reaction, 18 g of a diatomaceous earth filter aid was added in an amount of 3%, solid-liquid separation was carried out using a microporous filter plate (model: SS335), and the precipitate was collected for use.
4 dissolving the precipitate (immunoglobulin) collected in step (3) with 200mL of a 40mM sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution having a pH of 6.9, filtering the solution through a microfiltration filter plate (model: SS335) to remove diatomaceous earth and insoluble impurities, and further obtaining a clear solution containing purified equine immunoglobulin G, determining the protein content and adjusting the protein content to 6 mg/mL.
5, performing virus inactivation treatment, adding 0.5G/L medicinal rutin into the obtained clarified solution, enabling the clarified solution of the pure immune globulin G containing the rutin to flow through a helical quartz glass tube, and receiving UVC illumination with the intensity of 500-5000 mu w/cm2And (3) performing irradiation treatment for 120-1200 s.
6 directly loading the reaction solution after virus inactivation treatment for chromatographic separation (see FIG. 2)
(6) 10mL of an anionic chromatography medium (DEAE-sepharose Fast Flow) was loaded into an XK16/20 column;
(7) equilibrating the column with 20mM PBS buffer (pH7.0);
(8) injecting a chromatography sample loading solution containing horse immunoglobulin G into a chromatography column through an AKTA pure chromatograph, controlling the flow rate to be 1mL/min, monitoring a protein ultraviolet absorption peak, and collecting penetration liquid;
(9) after the sample loading is finished, continuously performing flow washing by using 20mM PBS buffer solution (pH7.0), and stopping collecting until the ultraviolet absorption peak of the protein is reduced to a baseline;
(10) after the flow washing is finished, changing the flow washing into 20mM PBS +1M NaCl buffer solution (pH7.0), eluting the hybrid protein combined with the chromatographic medium, and regenerating the chromatographic column;
7, performing ultrafiltration on the collected chromatography penetration peak solution by using an ultrafiltration membrane with the molecular weight cutoff of 50kDa, and replacing the solution with a horse immunoglobulin preservation buffer solution which is a 0.25mol/L glycine solution; the volume of the ultrafiltration exchange solution is 10 solution volumes, and the solution is concentrated until the content of the horse immunoglobulin is about 25 mg/mL; filtering and sterilizing by using a 0.2 mu m sterilizing filter membrane to obtain the high-purity horse immunoglobulin G solution.
Example 4 Virus inactivation Effect test
Canine immunoglobulin G was prepared according to the method of example 1, in which 10% of a virus solution (Pseudorabies virus, Pseudomonas virus, PRV, enveloped DNA virus; or Encephalomyocarditis virus, Encephalyomycarditis virus, EMCV, non-enveloped RNA virus) was mixed into a clarified solution of the purified immunoglobulin G.
In the virus inactivation treatment, the following treatments were respectively carried out on 6 samples (for reasons of space, screening experiments of conditions of various rutin types, light intensity and irradiation time are not shown): each sample treatment mode is carried out twice, and the inactivation effect on the pseudorabies virus and the encephalomyocarditis virus is verified respectively.
(sample 1) adding rutin injection 0.3g/L into the obtained clarified solution, mixing, placing into an inactivation instrument, and irradiating at 500 μ w/cm2And (6) irradiating for 1200 s.
(sample 2) adding rutin injection 0.3g/L into the obtained clarified solution, mixing, placing into an inactivation instrument, and irradiating with lightThe intensity is 5000 mu w/cm2And (4) irradiating for 120 s.
(sample 3) adding rutin injection 0.5g/L into the obtained clarified solution, mixing, placing into an inactivation instrument, and irradiating at 500 μ w/cm2And (6) irradiating for 1200 s.
(sample 4) the clear solution obtained was added with 0.5g/L rutin injection, mixed well and put into an inactivation apparatus with the illumination intensity of 5000. mu.w/cm2And (4) irradiating for 120 s.
(sample 5) adding rutin injection 0.8g/L into the obtained clarified solution, mixing, placing into an inactivation instrument, and irradiating at 500 μ w/cm2And (6) irradiating for 1200 s.
(sample 6) the obtained clear solution was added with 0.8g/L rutin injection, mixed well and put into an inactivation apparatus with the illumination intensity of 5000. mu.w/cm2And (4) irradiating for 120 s.
Calculation of Virus Titers (1) cells for detection (PK 15 cells for pseudorabies virus and BHK-21 cells for encephalomyocarditis virus) were set at 0.5 to 1.5 × 105Perml of cell culture plate with 96 wells, 100. mu.L per well; the culture plate was placed at 37 ℃ in 5% CO2Culturing in an incubator for 12-24 hours; (2) carrying out gradient continuous dilution on samples before and after treatment respectively, wherein the dilution times are 10 times; (3) add 10 to the cell culture plate-1To 10-10Serial gradient dilutions of virus, 100 μ L per well; (4) plates were incubated at 37 ℃ with 5% CO2Culturing in an incubator, observing cell lesions every day, and recording the number of lesion cell holes until cells in cell control holes can not maintain normal shapes; (5) calculating residual virus Titer (TCID) values of the samples before and after treatment according to the number of lesion holes with cytopathic effect of more than 50%50)。
The results of the tests on the samples of each group are shown in the following table:
the inactivation effect of the methods of samples 3 and 4 on the enveloped and non-enveloped viruses was further verified subsequently using Porcine parvovirus (PPV, non-enveloped DNA virus) and Vesicular stomatitis virus (VSV, enveloped RNA virus), which are common in animals. Results of virus-free detection after inactivation treatment were also obtained.
As can be seen, only 1 and 2 of the above-mentioned treatments detected virus residues, and the other treatments were effective in killing pseudorabies virus and encephalomyocarditis virus, and methods 3 and 4 are preferred from the viewpoint of preference.
The virus inactivation effect experiment shows that the method for preparing the high-purity immunoglobulin G has higher virus safety, and the application of the method has important application value for guaranteeing the safety of products.
Claims (1)
1. The use of medicinal rutin in the preparation of a UVC virus inactivation enhancer, wherein the UVC virus inactivation enhancer is used in the production process of high-purity immunoglobulin G; the UVC virus inactivation enhancer is used in a virus inactivation step in a high-purity immunoglobulin G production process; the production process for preparing the high-purity immunoglobulin G comprises the following steps:
(1) diluting animal serum raw material with normal saline until the protein concentration is less than or equal to 40 mg/mL;
(2) adding saturated ammonium sulfate solution at room temperature until the saturation reaches 30%, and centrifuging or filtering to remove impurity protein precipitate;
(3) collecting the supernatant or filtrate, adding saturated ammonium sulfate solution until saturation reaches 50%, centrifuging or filtering to collect immunoglobulin precipitate;
(4) dissolving immunoglobulin precipitate with 1-2 times of sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution, press-filtering with plate frame to remove insoluble impurities, further obtaining clear solution containing pure immunoglobulin G, and adjusting protein content to 5-10 mg/mL;
(5) virus inactivation: adding 0.5G/L medicinal rutin into the obtained clarified solution, allowing the clarified solution containing rutin-containing pure immunoglobulin G to flow through a helical quartz glass tube, and receiving UVC illumination with the intensity of 500-5000 μ w/cm2Performing irradiation treatment for 120-1200 s;
(6) ion exchange chromatography to obtain high-purity immunoglobulin G solution;
wherein the dissolving in the step (4) is performed using a 40mM sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution of pH 6.9; in the step (5), 0.5g/L of medicinal rutin is added into the clear solution obtained in the step (4); and treated with UVC light.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910770523.7A CN110872345B (en) | 2019-08-20 | 2019-08-20 | Preparation method of high-purity immunoglobulin G |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910770523.7A CN110872345B (en) | 2019-08-20 | 2019-08-20 | Preparation method of high-purity immunoglobulin G |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110872345A CN110872345A (en) | 2020-03-10 |
| CN110872345B true CN110872345B (en) | 2020-09-01 |
Family
ID=69716374
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910770523.7A Active CN110872345B (en) | 2019-08-20 | 2019-08-20 | Preparation method of high-purity immunoglobulin G |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110872345B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022071990A1 (en) * | 2020-04-10 | 2022-04-07 | Plasma Technologies, Llc | Compositions and methods for simplified high efficiency isolation of proteins |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2735986B2 (en) * | 1992-10-16 | 1998-04-02 | 株式会社セラリカ野田 | Antibodies to the product of the tumor suppressor gene WT |
| CN102512449A (en) * | 2012-01-09 | 2012-06-27 | 玉溪九洲生物技术有限责任公司 | Method for removing viruses in preparation of antitoxin and antiserum |
| CN105294860A (en) * | 2015-11-02 | 2016-02-03 | 张国强 | Preparation method and application of decayed tooth bacterium antibody |
| CN107496950B (en) * | 2017-08-18 | 2021-02-02 | 广州市众为生物技术有限公司 | Method for inactivating virus in protein solution by ultraviolet irradiation |
| CN108484760B (en) * | 2018-05-04 | 2021-08-31 | 中国人民解放军第三0二医院 | An anti-ricin immunoglobulin F (ab')2And method for preparing the same |
-
2019
- 2019-08-20 CN CN201910770523.7A patent/CN110872345B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN110872345A (en) | 2020-03-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4031833B2 (en) | Method for chromatographic removal of prions | |
| CN104672328B (en) | A kind of production method of Human Antithrombin Ⅲ | |
| CN111471103A (en) | Heterologous antibody of new coronavirus (2019-nCOV) and preparation method thereof | |
| KR19990036028A (en) | Industrial production method of Japanese encephalitis vaccine and vaccine by it | |
| TW201936628A (en) | Sterile chromatography and manufacturing processes | |
| CN107567496A (en) | Axenic purification method about virus | |
| CN110872345B (en) | Preparation method of high-purity immunoglobulin G | |
| CN105396129B (en) | Inactivated vaccine produced by poliomyelitis attenuated strain | |
| CN105601735B (en) | A kind of intravenous Giant cell human immunoglobulin and preparation method thereof | |
| CN112010968B (en) | Method for rapidly extracting blood plasma of patient with COVID-19 in convalescence stage for preparing immunoglobulin G | |
| CN101306200B (en) | Preparation method of hepatitis b human immunoglobulin for intravenous injection | |
| CN110241012B (en) | Production method and production module for upstream staged interception of biomacromolecules and application of production module in production | |
| CN112225799B (en) | Method for rapidly extracting blood plasma of COVID-19 patient in recovery period by automatic separation system | |
| CN105669860B (en) | A kind of hand-foot-and-mouth disease resistant human immunoglobulin and preparation method thereof | |
| CN108660119A (en) | A kind of purification process of Pseudorabies virus | |
| BR102013021499A2 (en) | essentially pure inactivated flavivirus virus preparation | |
| CN103937754A (en) | Porcine reproductive and respiratory syndrome virus (PPRSV) purification method | |
| CN105396128A (en) | Method for purifying poliomyelitis virus liquid | |
| CN110804578B (en) | Production method for removing IgG from bovine serum | |
| CN112500477B (en) | Method for rapidly extracting human immunoglobulin from blood plasma | |
| CN101497649B (en) | Production process of high fusion rate non-bacterial virus newborn bovine serum | |
| KR100900013B1 (en) | Method of purifying recombinant human erythropoietin from a fluid | |
| CN110904276A (en) | Application of bacteriophage in verification of virus removal process of biological product | |
| CN114573687B (en) | Storage method of human antithrombin III intermediate product | |
| KR102024342B1 (en) | Preparation methods for a novel generation of biological safe KLH Products used for cancer treatment, for the development of conjugated therapeutic vaccines and as challenging agents |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |