CN117860573A - Preparation of pharmaceutical compositions containing recombinant proteins - Google Patents
Preparation of pharmaceutical compositions containing recombinant proteins Download PDFInfo
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- CN117860573A CN117860573A CN202311698240.9A CN202311698240A CN117860573A CN 117860573 A CN117860573 A CN 117860573A CN 202311698240 A CN202311698240 A CN 202311698240A CN 117860573 A CN117860573 A CN 117860573A
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- 108010008281 Recombinant Fusion Proteins Proteins 0.000 title claims abstract description 52
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 title claims abstract description 52
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 6
- 238000011049 filling Methods 0.000 claims abstract description 162
- 230000002572 peristaltic effect Effects 0.000 claims abstract description 121
- 239000000203 mixture Substances 0.000 claims abstract description 67
- 238000001914 filtration Methods 0.000 claims abstract description 24
- 239000007788 liquid Substances 0.000 claims description 143
- 239000011550 stock solution Substances 0.000 claims description 59
- 238000003860 storage Methods 0.000 claims description 58
- 229930182555 Penicillin Natural products 0.000 claims description 50
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims description 50
- 229940049954 penicillin Drugs 0.000 claims description 50
- 238000000034 method Methods 0.000 claims description 34
- 239000011148 porous material Substances 0.000 claims description 33
- 238000004519 manufacturing process Methods 0.000 claims description 23
- 108010054265 Factor VIIa Proteins 0.000 claims description 20
- 238000010257 thawing Methods 0.000 claims description 19
- 108010039209 Blood Coagulation Factors Proteins 0.000 claims description 11
- 102000015081 Blood Coagulation Factors Human genes 0.000 claims description 11
- 239000003114 blood coagulation factor Substances 0.000 claims description 11
- 238000004806 packaging method and process Methods 0.000 claims description 8
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 1
- 230000008014 freezing Effects 0.000 claims 1
- 238000007710 freezing Methods 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 229910001220 stainless steel Inorganic materials 0.000 abstract description 21
- 239000010935 stainless steel Substances 0.000 abstract description 21
- 239000012905 visible particle Substances 0.000 abstract description 21
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 24
- 239000000741 silica gel Substances 0.000 description 24
- 229910002027 silica gel Inorganic materials 0.000 description 24
- 229940012414 factor viia Drugs 0.000 description 22
- 102000004169 proteins and genes Human genes 0.000 description 18
- 108090000623 proteins and genes Proteins 0.000 description 18
- 235000018102 proteins Nutrition 0.000 description 17
- 230000005484 gravity Effects 0.000 description 14
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- 238000007689 inspection Methods 0.000 description 11
- 229920001296 polysiloxane Polymers 0.000 description 11
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- 229940019700 blood coagulation factors Drugs 0.000 description 4
- 238000004891 communication Methods 0.000 description 4
- 238000012432 intermediate storage Methods 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 239000012906 subvisible particle Substances 0.000 description 4
- 206010053567 Coagulopathies Diseases 0.000 description 3
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- 230000035602 clotting Effects 0.000 description 3
- 230000006624 extrinsic pathway Effects 0.000 description 3
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- 230000004845 protein aggregation Effects 0.000 description 3
- 238000005096 rolling process Methods 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- 238000011179 visual inspection Methods 0.000 description 3
- 108010048049 Factor IXa Proteins 0.000 description 2
- 108010061932 Factor VIIIa Proteins 0.000 description 2
- 208000032843 Hemorrhage Diseases 0.000 description 2
- 108020004511 Recombinant DNA Proteins 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 230000015271 coagulation Effects 0.000 description 2
- 238000005345 coagulation Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 230000009851 immunogenic response Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 230000006623 intrinsic pathway Effects 0.000 description 2
- 229940112216 novoseven Drugs 0.000 description 2
- 108010013773 recombinant FVIIa Proteins 0.000 description 2
- 201000003542 Factor VIII deficiency Diseases 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 239000012270 PD-1 inhibitor Substances 0.000 description 1
- 239000012668 PD-1-inhibitor Substances 0.000 description 1
- 206010073391 Platelet dysfunction Diseases 0.000 description 1
- 108010000499 Thromboplastin Proteins 0.000 description 1
- 102000002262 Thromboplastin Human genes 0.000 description 1
- 208000013633 acquired hemophilia Diseases 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 229940125644 antibody drug Drugs 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 150000005829 chemical entities Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 208000019060 congenital factor VII deficiency Diseases 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 229960004222 factor ix Drugs 0.000 description 1
- 229940012413 factor vii Drugs 0.000 description 1
- 229960000301 factor viii Drugs 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000035931 haemagglutination Effects 0.000 description 1
- 208000009429 hemophilia B Diseases 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229960003301 nivolumab Drugs 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 229940121655 pd-1 inhibitor Drugs 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
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- 239000002243 precursor Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 230000006318 protein oxidation Effects 0.000 description 1
- 238000011110 re-filtration Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 231100000216 vascular lesion Toxicity 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
- A61J3/00—Devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D36/00—Filter circuits or combinations of filters with other separating devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B3/00—Packaging plastic material, semiliquids, liquids or mixed solids and liquids, in individual containers or receptacles, e.g. bags, sacks, boxes, cartons, cans, or jars
- B65B3/04—Methods of, or means for, filling the material into the containers or receptacles
- B65B3/10—Methods of, or means for, filling the material into the containers or receptacles by application of pressure to material
- B65B3/12—Methods of, or means for, filling the material into the containers or receptacles by application of pressure to material mechanically, e.g. by pistons or pumps
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Mechanical Engineering (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides a preparation method of a pharmaceutical composition containing recombinant protein, which is specifically improved on the basis of a disposable filtration and filling system, and a peristaltic pump (6) for filling is added with a back-end filter (7). The invention solves the problem that the recombinant protein composition filled by the traditional stainless steel tube tank system or the disposable filtering and filling system has visible particles.
Description
The application is a divisional application, the application date of the original application is 2019, 11, 13, 201911104057.5, and the name of the application is "preparation of a pharmaceutical composition containing recombinant proteins".
Technical Field
The invention relates to the field of proteins, in particular to a preparation method and production equipment of a pharmaceutical composition containing recombinant proteins, and particularly relates to a preparation method and production equipment of a composition containing antibodies or clotting polypeptides.
Background
Protein instability includes chemical instability and physical instability. Chemical instability involves the formation or destruction of covalent bonds, creating new chemical entities. In contrast, physical instability refers to the fact that the chemical composition is not changed, but the physical state of the protein is changed, such as denaturation, aggregation, precipitation, adsorption, etc. Among them, protein aggregation has become one of the most well-defined subjects in the field of protein stability.
Blood clotting phenomena involve a cascade of coagulation factors, indicated by numbers from I to XIII, which participate in coagulation in the reverse order of their numbering, except for factors XIII, which participate in the last step of the clotting reaction, each factor being present in the form of an inactive precursor and in the activated form indicated by letter a. Blood coagulation involves two pathways: intrinsic and extrinsic pathways. Wherein the intrinsic pathway comprises factors present in the blood circulation and the clotting process starts from within the blood vessel. The extrinsic pathway involves coagulation factors that are not entirely present in the blood, but also tissue factors that are released in vascular lesions. The extrinsic pathway involves the intervention of factor vii (vii), activated factor viia being one of the factors involved in the hemagglutination mechanism, with a molecular weight of about 50 kDa. Fviia binds tissue factor and initiates subsequent activation of the coagulation factor, resulting in a coagulation cascade. Fviia can act locally even in the absence of factor viii or ix.
NovoSeven RT developed by Danenode and Norde is recombinant human factor VIIa, similar in structure to human plasma-derived factor VIIa. It has been approved for bleeding event and perioperative management in haemophilia a/B patients, congenital factor VII deficiency patients and platelet dysfunction patients, and for bleeding event and perioperative management in acquired haemophilia patients.
As a protein medicine, the FVIIa has poor stability in a solution state and is easy to generate physical and chemical changes under the action of external factors, thereby influencing the effectiveness and safety of the medicine. NovoSeven RT, developed and produced by Norand Norde, is a method of maintaining long-term stability of the product by lyophilization. However, during the production of the preparation, the interfacial stability of the contact between the tube and the feed liquid, the generation and breaking of bubbles, the shearing force during the transmission, and the like may cause aggregation of proteins, generate sub-visible and visible particles, and increase the possibility of adverse immunogenic reactions during the treatment.
IgG4 antibody drugs represented by the PD-1 inhibitor Nivolumab (Na Wu Liyou antibody) also have similar problems of protein aggregation stability during preparation production. This is probably because IgG4 antibodies are structurally distinct from other subtypes, resulting in faster aggregation rates under agitation and high temperature conditions and also in the formation of more sub-visible particles when contacted with stainless steel reservoirs, EVA-based reservoirs, etc.
The traditional workshop production, preparation and filtration adopts a stainless steel pipe tank system, stock solution is mixed in a stainless steel tank, the stock solution is pressurized by compressed air and is sterilized and filtered by a two-stage 0.22 mu m pore size filter and is pressed into a stainless steel liquid storage tank, and the tank bodies are connected by using a stainless steel pipe. The sterilized and filtered feed liquid is stored in a stainless steel tank, compressed air is adopted to convey the feed liquid to a stainless steel buffer tank in a pressurizing mode, and then peristaltic pump is used for filling. With this system, metal ions in the stainless steel tube can catalyze protein oxidation, making the protein unstable; the solid-liquid interface effect of the feed liquid contacted with the tube tank can also cause unstable protein and aggregation; the use of compressed air for pressurization can lead to a large amount of generation of bubbles, the generation and the breaking of the bubbles can lead to the great increase of gas-liquid interfaces, and the action of the gas-liquid interfaces can also damage the protein structure to cause protein aggregation. When the traditional technology is used for production, once the protein in the filtered feed liquid is aggregated to generate sub-visible and visible particles, the sub-visible and visible particles are packaged in penicillin bottles and then are freeze-dried, so that the particles in the freeze-dried finished preparation product can be increased. The microparticles are injected into the human body, may generate an immunogenic reaction, even block blood vessels, and may be harmful to the user.
At present, a disposable filtering and filling system is increasingly used in the production of biological medicines, a liquid storage bag made of high-molecular polymer materials is used for replacing a stainless steel tank, a silica gel hose is used for replacing a stainless steel pipeline, and a peristaltic pump is used for conveying feed liquid to replace compressed air. Compared with the traditional technology, the technology reduces the risk of stainless steel influence and reduces the generation of bubbles. However, for some proteins with poor stability and greater influence of hydrophobic forces (e.g. fviia, igG4 antibodies), aggregation may still occur when the proteins are contacted with a liquid storage bag with a hydrophobic inner surface, and such a liquid may be directly filled into a penicillin bottle for lyophilization, which may result in a large number of visible particles in the finished product.
The research shows that the raw liquid of the FVIIa and the IgG4 antibody has few particles after being subjected to sterilization and filtration, meets the Chinese pharmacopoeia standard, but the particles of the feed liquid are obviously increased after the raw liquid passes through a stainless steel liquid storage tube (or liquid storage bag) and is filled into a penicillin bottle. There is an urgent need for a new preparation production device and process that can reduce the particle content of proteins with poor stability such as fviia and IgG4 antibodies after filling, and improve the quality and safety thereof.
In the production of recombinant protein preparation, peristaltic pump for filling is directly connected with filling needle. We have tried to add a filter between the reservoir bag (4) and the dispenser (5) or between the dispenser (5) and the peristaltic pump (6) for filling to achieve re-filtration before filling, however neither of the above improvements fundamentally solves the visible particulate problem of the product after filling.
Disclosure of Invention
The invention aims at providing a production device and a preparation method of a recombinant protein composition.
In one aspect, the present invention provides an apparatus for producing a recombinant protein composition. In particular, the invention is an improvement on the basis of a disposable filtration and filling system: a rear end filter (7) is additionally arranged behind a peristaltic pump (6) for filling connected behind the liquid separator (5), namely, each peristaltic pump (6) for filling is respectively connected with a rear end filter (7) and a filling needle head (8) in sequence.
In some aspects, the production apparatus includes the following elements: stock solution storage bags or stock solution freeze thawing bags, front peristaltic pumps, filters, middle liquid storage bags, liquid separators, peristaltic pumps for filling, filling needles and/or the like. The raw liquid storage bag or the raw liquid freeze thawing bag is used for storing raw liquid of the recombinant protein composition, the raw liquid storage bag is made of high polymer materials, the peristaltic pump is used for providing filtering or filling power, the filter is used for filtering feed liquid of the recombinant protein composition, the middle liquid storage bag is used for storing the filtered feed liquid of the recombinant protein composition, the liquid separator is used for separating the recombinant protein composition or the recombinant protein medicine, and the filling needle is used for filling the feed liquid of the recombinant protein composition into penicillin bottles.
In some embodiments, the elements are connected by pipes, for example, hard or soft pipes, preferably by silicone hoses.
In some aspects, the production facility includes an isolator that is within a sterile environment.
In some aspects, the production facility includes components in communication, specifically, a stock solution reservoir or a stock solution freeze-thaw bag, a front end peristaltic pump, 2 or more front end filters, an intermediate reservoir, a dispenser, a peristaltic pump for filling, and/or a filling needle, etc. In some schemes, the stock solution liquid storage bag or the stock solution freeze thawing bag is connected with a front peristaltic pump, 2 front filters and a middle liquid storage bag in sequence, the middle liquid storage bag is connected with a later liquid separator, and the liquid separator is connected with a peristaltic pump for filling and a filling needle in sequence. In some embodiments, the filling needle is followed by a penicillin bottle to effect filling of the protein composition liquor. In one embodiment, the dispenser is positioned below the intermediate reservoir to create a height differential to effect gravity transfer of the feed liquid in the reservoir to the dispenser. In some embodiments, 1 front end filter, intermediate reservoir bag, dispenser, peristaltic pump for filling, and/or filling needle are located in the separator.
In some aspects, the production facility includes components in communication, specifically, 1 or more stock solution storage bags or stock solution freeze thawing bags, 1 or more front end peristaltic pumps, 2 or more front end filters, 1 or more intermediate storage bags, 1 or more dispensers, 1 or more peristaltic pumps for filling, and/or 1 or more filling needles, etc.
In some schemes, each production device at least comprises 1 stock solution storage bag or stock solution freeze thawing bag, 1 front end peristaltic pump, 2 front end filters connected in series, 1 middle storage bag, 1 knockout, n peristaltic pumps for filling in parallel, and each peristaltic pump for filling is respectively connected with one filling needle. Wherein n is an integer from 1 to 24, and may be, for example, 1,2,3,4,5,6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24.
In some embodiments, the stock solution storage bag or the stock solution freeze-thaw bag, the front end peristaltic pump, the front end filter, the middle liquid storage bag, the liquid separator, the peristaltic pump for filling, and/or the filling needle may further be inserted or added between each two elements or between two elements, wherein the added elements are in communication with the stock solution storage bag or the stock solution freeze-thaw bag, the front end peristaltic pump, the front end filter, the middle liquid storage bag, the liquid separator, the peristaltic pump for filling, or the filling needle through pipes and/or elements. The pipe can be a hard pipe or a soft pipe, preferably a silica gel hose. The inserted or added other element may be a filter.
In some embodiments, a stock solution reservoir or a stock solution freeze-thaw bag, a front peristaltic pump, a front filter, an intermediate reservoir, a dispenser, a peristaltic pump for filling, and/or a filling needle may be added or inserted between each two elements, or between the two elements, and the filter is connected to the elements at the two ends thereof by a pipe. The pipe can be a hard pipe or a soft pipe, preferably a silica gel hose.
In some embodiments, the stock solution storage bag or stock solution freeze-thaw bag, front end peristaltic pump, front end filter in series, intermediate storage bag, dispenser, peristaltic pump for filling, and/or filling needle are sequentially connected in series. In some embodiments, the stock solution storage bag or stock solution freeze-thaw bag, the front end peristaltic pump, the front end filter, the intermediate storage bag, the dispenser, the peristaltic pump for filling, and/or the additional element may be inserted or added between or between each two elements of the filling needle, wherein the added element is in communication with the stock solution storage bag or stock solution freeze-thaw bag, the front end peristaltic pump, the front end filter, the intermediate storage bag, the dispenser, the peristaltic pump for filling, or the filling needle via a conduit or element. In some embodiments, the elements are connected by pipes, for example, hard or soft pipes, preferably by silicone hoses. In some embodiments, the stock solution storage bag or the stock solution freeze-thawing bag, the front peristaltic pump, the front filter, the middle storage bag, the liquid separator, the peristaltic pump for filling, and/or the filling needle can be further added or inserted between each two elements or at two ends, and the filter is connected with the elements at two ends thereof through a pipeline. In some embodiments, the filling needle fills the protein composition to a penicillin bottle. In some embodiments, the 1 front-end filter, the intermediate reservoir, the dispenser, the peristaltic pump for filling, the filling needle and/or the penicillin bottle are located in one isolator.
In some embodiments, each set of production equipment is formed by connecting 1 stock solution storage bag or stock solution freeze thawing bag, 1 front end peristaltic pump, 2 front end filters connected in series, 1 middle storage bag, 1 liquid separator, 1-12 peristaltic pumps for filling, and/or 1-12 filling needles. In some specific embodiments, the stock solution storage bag or the stock solution freeze thawing bag, the front peristaltic pump, the 2 front filters and the middle storage bag are sequentially connected in series, the liquid separator is connected in series with 1-12 peristaltic pumps for filling, the peristaltic pumps for filling are connected in parallel, and each peristaltic pump for filling is sequentially connected in series with 1 back filter and 1 filling needle. In some embodiments, the elements are connected by pipes, for example, hard or soft pipes, preferably by silicone hoses. In some embodiments, the filling needle fills the protein composition into a penicillin bottle (9). In some embodiments, the 1 front-end filter, the intermediate reservoir, the dispenser, the peristaltic pump for filling, the back-end filter, the filling needle and/or the penicillin bottle are located in one isolator.
In some embodiments, the front end filter pore size between the front end peristaltic pump and the reservoir bag is 0.1 μm to 1 μm, in some embodiments the filter pore size is 0.1 μm,0.22 μm,0.45 μm, in one particular embodiment the filter pore size is 0.22 μm.
In some embodiments, the back end filter pore size between the peristaltic pump and the filling needle is 0.1 μm to 1 μm. In some embodiments, the filter pore size is 0.1 μm,0.22 μm,0.45 μm. In a particular embodiment, the filter pore size is 0.22 μm.
In some embodiments, the back end filter between the peristaltic pump and the filling needle is a combination filter of pore sizes, wherein each pore size is 0.1 μm to 1 μm, respectively. In some embodiments, each of the combined pore sizes is 0.1 μm,0.22 μm,0.45 μm, respectively, and in some embodiments, at least one of the combined pore sizes is 0.22 μm.
In some aspects, the invention provides an apparatus for producing a recombinant protein composition, the apparatus comprising a disposable filtration and filling system. In some embodiments, the disposable filtration filling system comprises a disposable liquid storage bag or a disposable freeze-thaw bag. In some embodiments, the liquid storage bag or the freeze thawing bag is used for storing the recombinant protein composition and is made of high molecular polymer materials. In some embodiments, the disposable filtration filling system comprises a disposable tubing, preferably a silicone hose, connecting each adjacent element. In some embodiments, the disposable filtration filling system further comprises all other necessary elements and/or tubing to complete the preparation of the recombinant protein composition.
In some embodiments, the recombinant protein is selected from the group consisting of factor, preferably recombinant human factor, more preferably recombinant human factor VIIa, factor VIIIa, or factor IXa, most preferably recombinant human factor VIIa.
In some embodiments, the recombinant protein is an antibody, preferably an IgG antibody, more preferably an IgG4 antibody, most preferably a nano Wu Liyou mab.
In some aspects, the invention provides an apparatus for producing a recombinant protein composition comprising the following elements: 1 stock solution bag or freeze thawing bag (1) for storing recombinant protein composition stock solution, which is connected with 1 front end peristaltic pump (2), 2 front end filters ((3) and (3')) with 0.22 μm in series, 1 middle stock solution bag (4), 1 knockout (5), 6 peristaltic pumps for filling (6), 6 rear end filters (7) with 0.22 μm in pore size, 6 filling needles (8), 6 penicillin bottles (9) in sequence. Wherein the 1 front-end filter, the middle liquid storage bag, the liquid distributor, the 6 peristaltic pumps for filling, the 6 rear-end filters and the 6 penicillin bottles are positioned in one isolator (10).
In one aspect, the invention provides a method of preparing a recombinant protein composition. In another aspect, the invention provides a method of filling a recombinant protein composition or recombinant protein medicament. In some embodiments, the invention is an improvement over the disposable filtration-fill preparation method.
In one aspect, the present invention provides methods of preparing, filling, packaging, and/or producing recombinant protein compositions. In some embodiments, the method comprises filling the recombinant protein composition. In some embodiments, the filled recombinant protein composition comprises: i) Filtering the recombinant protein composition using a filter, ii) filling the composition into penicillin bottles using a filling needle. In some embodiments, the filter is a back end filter that is directly connected to the filling needle by tubing. In some embodiments, the preparation of the recombinant protein composition is performed using the apparatus of the present invention. In some embodiments, the recombinant protein is selected from a factor or IgG4 antibody, preferably a recombinant human factor VIIa or nal Wu Liyou mab.
In some embodiments, the present invention provides methods of preparing a recombinant protein composition, the methods comprising filtering the recombinant protein composition prior to filling. In some embodiments, the protocol comprises dispensing the recombinant protein composition or recombinant protein drug using a dispenser. Wherein, each peristaltic pump for filling that connects behind the knockout is connected a rear end filter respectively afterwards, makes peristaltic pump for filling connect gradually a filter, a filling syringe needle and a penicillin bottle respectively.
In some embodiments, the methods of preparing a recombinant protein composition provided herein include the steps of: 1. filtering the recombinant protein composition stock solution in a liquid storage bag or a freeze thawing bag (1) by using a front peristaltic pump (2) to provide filtering power, and performing secondary filtration through 2 front filters ((3) and (3')) to an intermediate liquid storage bag (4) in an isolator; 2. the liquid separator (5) is arranged below the middle liquid storage bag (4) to form a height difference, and the liquid in the liquid storage bag is conveyed into the liquid separator by utilizing the gravity effect. The recombinant protein composition feed liquid in the knockout sequentially passes through a peristaltic pump (6) for filling, a rear end filter (7) and a filling needle head (8) and enters a penicillin bottle (9). Wherein 1 front filter, middle liquid storage bag, liquid distributor, peristaltic pump for filling, back filter, filling needle and/or penicillin bottle are/is located in one isolator (10).
In some embodiments, the number of peristaltic pumps and/or needles for filling connected after the dispenser is n, n being an integer from 1 to 24, which may be, for example, 1,2,3,4,5,6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24. In some embodiments, the n peristaltic pumps and/or the filling needles are connected in parallel. In some embodiments, n is an integer from 1 to 12, which may be, for example, 1,2,3,4,5,6,7,8,9, 10, 11, 12. In a particular embodiment, n=6.
In some embodiments, the front end filter pore size between the front end peristaltic pump and the reservoir bag is 0.1 μm to 1 μm, in some embodiments, the front end filter pore size is 0.1 μm,0.22 μm,0.45 μm, in one particular embodiment, the front end filter pore size is 0.22 μm.
In some embodiments, the back end filter pore size between the peristaltic pump and the filling needle is 0.1 μm to 1 μm. In some embodiments, the filter pore size is 0.1 μm,0.22 μm,0.45 μm. In a particular embodiment, the filter pore size is 0.22 μm.
In some embodiments, the back end filter between the peristaltic pump and the filling needle is a combination filter of pore sizes, wherein each pore size is 0.1 μm to 1 μm, respectively. In some embodiments, each of the combined pore sizes is 0.1 μm,0.22 μm,0.45 μm, respectively, and in some embodiments, at least one of the combined pore sizes is 0.22 μm.
In some embodiments, the recombinant protein is selected from the group consisting of factor, preferably recombinant human factor, more preferably recombinant human factor VIIa, factor VIIIa, or factor IXa, most preferably recombinant human factor VIIa.
In some embodiments, the recombinant protein is an antibody, preferably an IgG antibody, more preferably an IgG4 antibody, most preferably a nano Wu Liyou mab.
The present invention provides a device for the production of a composition comprising recombinant human factor VIIa or IgG4 antibodies comprising the following elements:
1. a reservoir for storing a stock solution of a composition comprising recombinant human factor VIIa or a freeze-thaw bag of a stock solution of a composition comprising IgG4 antibodies, which reservoir or freeze-thaw bag is connected in sequence to a peristaltic pump (which may be referred to as a front end peristaltic pump), two filters with a pore size of 0.22 μm in series (which may be referred to as a front end filter) and an intermediate reservoir located within the isolator, the individual components being connected by means of a silicone hose.
2. A knockout passes through the silica gel hose to be connected in the below of middle stock solution bag, forms the difference in height to realize utilizing the gravity effect to transmit the feed liquid in the stock solution bag to the knockout in.
3. Six peristaltic pumps for filling are connected through a silica gel hose after the liquid separator, then each peristaltic pump is respectively connected with a filter with the aperture of 0.22 mu m and a filling needle, and a penicillin bottle is arranged after the filling needle, so that feed liquid in the liquid separator can sequentially pass through the peristaltic pumps, the filter and the filling needle and enter the penicillin bottle.
The present invention provides an apparatus for the production of a composition comprising recombinant human factor VIIa comprising the following elements:
1. a reservoir for storing a stock solution of a composition comprising recombinant human factor viia, said reservoir being connected in sequence to a peristaltic pump (which may be referred to as a front end peristaltic pump), two filters of pore size 0.22 μm in series (which may be referred to as front end filters) and an intermediate reservoir within the separator, the individual components being connected by means of silicone hoses.
2. A knockout passes through the silica gel hose to be connected in the below of middle stock solution bag, forms the difference in height to realize utilizing the gravity effect to transmit the feed liquid in the stock solution bag to the knockout in.
3. Six peristaltic pumps for filling are connected through a silica gel hose after the liquid separator, then each peristaltic pump is respectively connected with a filter with the aperture of 0.22 mu m and a filling needle, and a penicillin bottle is arranged after the filling needle, so that feed liquid in the liquid separator can sequentially pass through the peristaltic pumps, the filter and the filling needle and enter the penicillin bottle.
The invention provides a production device of a composition containing an IgG4 antibody, which comprises the following elements:
1. the freeze-thawing bag for storing the stock solution of the composition containing the IgG4 antibody is connected with a peristaltic pump (which can be called a front peristaltic pump), two filters with the pore diameter of 0.22 mu m (called front filters) connected in series and an intermediate liquid storage bag positioned in the isolator in sequence, and all the components are connected by using a silica gel hose respectively.
2. A knockout passes through the silica gel hose to be connected in the below of middle stock solution bag, forms the difference in height to realize utilizing the gravity effect to transmit the feed liquid in the stock solution bag to the knockout in.
3. Six peristaltic pumps for filling are connected through a silica gel hose after the liquid separator, then each peristaltic pump is respectively connected with a filter with the aperture of 0.22 mu m and a filling needle, and a penicillin bottle is arranged after the filling needle, so that feed liquid in the liquid separator can sequentially pass through the peristaltic pumps, the filter and the filling needle and enter the penicillin bottle.
The invention provides a preparation method of a composition containing recombinant human coagulation factor VIIa or IgG4 antibody, which comprises the following steps:
1. the stock solution of the recombinant human factor VIIa composition in the stock solution bag or the stock solution of the IgG4 antibody composition in the freeze thawing bag is filtered by a peristaltic pump (which may be called a front peristaltic pump) to obtain a filtered solution, and the filtered solution is filtered by two filters with 0.22 μm diameter connected in series to an intermediate stock solution bag in an isolator, and the components are connected by a silica gel hose.
2. The liquid distributor is arranged below the middle liquid storage bag to form a height difference, and the liquid in the liquid storage bag is transmitted to the liquid distributor by utilizing the gravity effect, and all the components are connected by using a silica gel hose.
3. And after the liquid separator, six peristaltic pumps for filling are connected by using a silica gel hose, each peristaltic pump is sequentially connected with a filter with the aperture of 0.22 mu m, a filling needle head and a penicillin bottle, and the liquid in the liquid separator sequentially passes through the peristaltic pumps, the filter and the filling needle head and enters the penicillin bottle.
The invention provides a preparation method of a composition containing recombinant human coagulation factor VIIa, which comprises the following steps:
1. the stock solution of the recombinant human factor VIIa composition in the reservoir was filtered using peristaltic pump (which may be referred to as front end peristaltic pump) and filtered through two series connected filters of 0.22 μm diameter into a middle reservoir located in the isolator using a silicone hose connection between the components.
2. The liquid distributor is arranged below the middle liquid storage bag to form a height difference, and the liquid in the liquid storage bag is transmitted to the liquid distributor by utilizing the gravity effect, and all the components are connected by using a silica gel hose.
3. And after the liquid separator, six peristaltic pumps for filling are connected by using a silica gel hose, each peristaltic pump is sequentially connected with a filter with the aperture of 0.22 mu m, a filling needle head and a penicillin bottle, and the liquid in the liquid separator sequentially passes through the peristaltic pumps, the filter and the filling needle head and enters the penicillin bottle.
The invention provides a preparation method of a composition containing an IgG4 antibody, which comprises the following steps:
1. the stock solution of the IgG4 antibody composition contained in the freeze-thawing bag was filtered by two stages through two filters with a pore size of 0.22 μm connected in series into an intermediate reservoir bag located in the separator using a peristaltic pump (which may be referred to as a front peristaltic pump), and the components were connected by a silicone hose.
2. The liquid distributor is arranged below the middle liquid storage bag to form a height difference, and the liquid in the liquid storage bag is transmitted to the liquid distributor by utilizing the gravity effect, and all the components are connected by using a silica gel hose.
3. And after the liquid separator, six peristaltic pumps for filling are connected by using a silica gel hose, each peristaltic pump is sequentially connected with a filter with the aperture of 0.22 mu m, a filling needle head and a penicillin bottle, and the liquid in the liquid separator sequentially passes through the peristaltic pumps, the filter and the filling needle head and enters the penicillin bottle.
The invention solves the problem that the recombined protein composition feed liquid filled by the traditional stainless steel tube tank system or the disposable filtering and filling system may have visible particles. Surprisingly, we have found that unexpected technical results are achieved using the production apparatus and the preparation method of the present invention. Specifically, when the traditional stainless steel tube tank system is used, the filled product has a large amount of visible particles, when the disposable filtering and filling system is used, the filled product also has a small amount or very small amount of visible particles, and when the method and the device are used, the filled product has no visible particles, so that the requirements of Chinese pharmacopoeia are met.
For the purposes of this application, the following terms, as used in the specification and claims, shall have the following meanings, unless otherwise indicated.
"recombinant protein": refers to a protein obtained by applying recombinant DNA technology or recombinant RNA technology.
"coagulation factor": refers to various protein components involved in the blood coagulation process, including blood coagulation factors I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII.
"recombinant human coagulation factor": is a human coagulation factor produced by recombinant DNA technology.
"antibody": in its broadest sense, in particular intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g. bispecific antibodies) formed from at least two intact antibodies, and antibody fragments are contemplated, provided they possess the desired biological activity.
IgG antibodies: igs are classified into five classes, igG, igA, igM, igD and IgE, depending on the heavy chain, where an IgG antibody refers to an Ig whose heavy chain is a gamma chain. Wherein IgG is further divided into four subclasses, igG1-IgG 4.
Monoclonal antibody: also called monoclonal antibodies, refer to specific antibodies against a single epitope.
Drawings
Fig. 1 is a schematic view of a production apparatus of the present invention.
Detailed Description
Example 1
A composition for preparing recombinant human factor viia using a stainless steel tube tank system comprising:
(1) 10L of the recombinant human factor VIIa stock solution in a reservoir bag was filtered through 2 filters of 0.22 μm pore size using a stainless steel tube tank system, powered with compressed air, to a stainless steel reservoir tank and transferred to a buffer tank in an isolator.
(2) The feed liquid in the buffer tank is transferred to the liquid separator under the action of gravity.
(3) The liquid distributor is connected with 6 peristaltic pumps, the peristaltic pumps are started, and the feed liquid passes through the peristaltic pumps and the filling needle heads, enters the penicillin bottles and is filled according to 1 mg/bottle. The filling amount was checked during filling, and the fluctuation range of the filling amount was ±2%.
(4) And (3) plugging the penicillin bottle, transmitting the penicillin bottle into a freeze dryer for freeze-drying, rolling the penicillin bottle out of the box after freeze-drying, performing visual inspection, and packaging to obtain a finished product.
(5) Taking 5 bottles of finished products, respectively slowly injecting 1ml of a special solvent for recombinant human blood coagulation factor VIIa into a penicillin bottle, slowly rotating to completely dissolve, and checking visible foreign matters according to the Lamp inspection method of the national formulary (2015 edition) general rule 0904 visible foreign matter inspection method.
The specific operation of the light inspection method is as follows: taking a specified amount of the test substance, removing the container label, wiping the outer wall of the container, placing the test substance at the edge of the light shielding plate, holding the neck of the container at a clear vision distance (the clear observation distance from the test substance to human eyes is usually 25 cm), lightly rotating and turning the container (but avoiding generating bubbles), suspending the visible foreign matters possibly existing in the liquid medicine, visually inspecting in black and white backgrounds respectively, and repeatedly observing, wherein the total inspection time limit is 20 seconds. When a large number of bubbles in the sample solution affect observation, the sample solution needs to be kept for enough time until the bubbles disappear, and then the sample solution is checked. For the injections, 20 samples (bottles) were taken and examined as described above, unless otherwise specified. For the sterile preparation for injection, 5 test pieces (bottles) were taken, and after the powder was completely dissolved by a suitable solvent and a suitable method, the test pieces were examined as described above.
And (3) result judgment: the tested formulations were semi-quantitatively classified according to the contamination of the visible particles:
a = no visible particles
B=very small amount of visible particles (3 or less of fine visible foreign matters in each bottle)
C=a small amount of visible particles (more than 3 and 10 or less of fine visible foreign matters in each (bottle))
D=a large number of visible particles (more than 10 and 50 or less of fine visible foreign matters per bottle)
E=very large number of visible particles (more than 50 fine visible foreign matter in each (bottle))
Note that: the fine visible foreign matter is pointed object, short fiber and lump with size below 2mm, or semitransparent small recombinant protein floccule or recombinant protein granule with size less than about 1mm in biological product
The condition of the finished product particles after 5 bottles are grade D, and a large number of visible particles are not in accordance with the requirements of Chinese pharmacopoeia (2015 edition).
Example 2
A composition for preparing recombinant human factor viia using a disposable filtration and fill system comprising:
(1) 10L of the recombinant human factor VIIa stock solution in the reservoir was filtered through 2 filters of 0.22 μm pore size into a 20L intermediate reservoir placed in an isolator using peristaltic pump and connected in between using a silicone hose.
(2) The liquid storage bag in the isolator is connected with the liquid separator through a silica gel hose, and the liquid in the liquid storage bag is transmitted to the liquid separator under the action of gravity.
(3) The liquid distributor is connected with 6 peristaltic pumps, the peristaltic pumps are started, and the feed liquid passes through the peristaltic pumps and the filling needle heads, enters the penicillin bottles and is filled according to 1 mg/bottle. The filling amount was checked during filling, and the fluctuation range of the filling amount was ±2%.
(4) And (3) plugging the penicillin bottle, transmitting the penicillin bottle into a freeze dryer for freeze-drying, rolling the penicillin bottle out of the box after freeze-drying, performing visual inspection, and packaging to obtain a finished product.
(5) Taking 5 bottles of finished products, respectively slowly injecting 1ml of a special solvent for recombinant human blood coagulation factor VIIa into a penicillin bottle, slowly rotating to completely dissolve, and checking visible foreign matters according to the Lamp inspection method of the national formulary (2015 edition) general rule 0904 visible foreign matter inspection method.
The condition of the finished product particles after 5 bottles are C-grade, and a small amount of visible particles are not in accordance with the requirements of Chinese pharmacopoeia (2015 edition).
Example 3
A composition for preparing recombinant human factor viia using the protocol of the present invention comprises:
(1) 10L of the recombinant human factor VIIa stock solution contained in the reservoir bag was filtered by a peristaltic pump with a secondary filtration through 2 filters of 0.22 μm in diameter into a 20L intermediate reservoir bag placed in an isolator with a silicone hose connection in between.
(2) The liquid distributor is arranged below the middle liquid storage bag to form a height difference, and the liquid in the liquid storage bag is transmitted to the liquid distributor by utilizing the gravity effect, and the middle part is connected by using a silica gel hose.
(3) And after the liquid separator, six peristaltic pumps for filling are connected by using a silica gel hose, each peristaltic pump is sequentially connected with a filter with the aperture of 0.22 mu m, a filling needle head and a penicillin bottle, and the liquid in the liquid separator sequentially passes through the peristaltic pumps, the filter and the filling needle head and enters the penicillin bottle. Filling was performed at 1 mg/bottle. The filling amount was checked during filling, and the fluctuation range of the filling amount was ±2%.
(4) And (3) plugging each penicillin bottle, transmitting to a freeze dryer for freeze drying, rolling a cover out of the box after freeze drying, performing visual inspection, and packaging to obtain a finished product.
(5) Taking 5 bottles of finished products, respectively slowly injecting 1ml of a special solvent for recombinant human blood coagulation factor VIIa into a penicillin bottle, slowly rotating to completely dissolve, and checking visible foreign matters according to the Lamp inspection method of the national formulary (2015 edition) general rule 0904 visible foreign matter inspection method.
The condition of the finished product particles after 5 bottles are grade A, no visible particles exist, and the requirements of Chinese pharmacopoeia (2015 edition) are met.
Example 4
Preparation of recombinant human factor VIIa compositions using the following production equipment
The modification of the disposable filtration and filling system described in example 2 was performed by adding a filter between the intermediate reservoir bag and the dispenser or between the dispenser and the peristaltic pump for filling.
The recombinant human factor VIIa composition was filtered and filled using the apparatus, again at 1 mg/bottle, into a penicillin bottle. The filling amount was checked during filling, and the fluctuation range of the filling amount was ±2%. Then, the following steps of plugging, freeze-drying, checking for visible foreign matters and the like are carried out.
As a result, it was found that the composition obtained by filling with these two apparatuses contained a small amount of visible fine particles, and could not meet the requirements of the Chinese pharmacopoeia (2015 edition).
Example 5
A composition for preparing a nano Wu Liyou antibody using a stainless steel tube tank system comprising:
(1) 10L of the Wu Liyou antibody stock solution contained in a freeze-thaw bag was filtered through 2 filters of 0.22 μm pore size to a stainless steel reservoir tank using a stainless steel tube tank system powered with compressed air and transferred to an isolator buffer tank.
(2) The feed liquid in the buffer tank is transferred to the liquid separator under the action of gravity.
(3) The liquid distributor is connected with 6 peristaltic pumps, the peristaltic pumps are started, and the feed liquid passes through the peristaltic pumps and the filling needle heads, enters the penicillin bottles and is filled according to 4 ml/bottle. The filling amount was checked during filling, and the fluctuation range of the filling amount was ±2%.
(4) And (5) plugging the penicillin bottle, capping, checking with eyes, and packaging to obtain a finished product.
(5) Taking 20 bottles of finished products of the preparation, and checking visible foreign matters according to the lamp inspection method of the general rule 0904 visible foreign matter checking method of Chinese pharmacopoeia (2015 edition).
The 20 bottles of the finished particles are grade D, have a large number of visible particles and do not meet the requirements of Chinese pharmacopoeia (2015 edition).
Example 6
A composition for preparing a nano Wu Liyou antibody using a disposable filtration and filling system comprising:
(1) The 10L of the Wu Liyou antibody stock solution contained in the freeze thawing bag was powered by peristaltic pump and filtered through 2 filters with 0.22 μm pore size into a 20L intermediate reservoir placed in an isolator, the middle was connected by a silicone hose.
(2) The middle liquid storage bag in the isolator is connected with the liquid separator through a silica gel hose, and the liquid in the liquid storage bag is transmitted to the liquid separator under the action of gravity.
(3) The liquid distributor is connected with 6 peristaltic pumps, the peristaltic pumps are started, and the feed liquid passes through the peristaltic pumps and the filling needle heads, enters the penicillin bottles and is filled according to 4 ml/bottle. The filling amount was checked during filling, and the fluctuation range of the filling amount was ±2%.
(4) And (5) plugging the penicillin bottle, capping, checking with eyes, and packaging to obtain a finished product.
(5) Taking 20 bottles of finished products of the preparation, and checking visible foreign matters according to the lamp inspection method of the general rule 0904 visible foreign matter checking method of Chinese pharmacopoeia (2015 edition).
The 20 bottles of the finished particles are all grade B, have a very small amount of visible particles and meet the requirements of Chinese pharmacopoeia (2015 edition).
Example 7
A composition for preparing a nano Wu Liyou antibody using the protocol of the present invention, comprising:
(1) 10L of the crude antibody solution containing Wu Liyou antibody in a freeze thawing bag is filtered by a peristaltic pump to obtain a filtered solution, and the filtered solution is filtered by 2 filters with the pore diameter of 0.22 mu m to a 20L middle liquid storage bag arranged in an isolator, and the middle liquid storage bag is connected by a silica gel hose.
(2) The liquid distributor is arranged below the middle liquid storage bag to form a height difference, and the liquid in the liquid storage bag is transmitted to the liquid distributor by utilizing the gravity effect, and the middle part is connected by using a silica gel hose.
(3) And after the liquid separator, six peristaltic pumps for filling are connected by using a silica gel hose, each peristaltic pump is sequentially connected with a filter with the aperture of 0.22 mu m, a filling needle head and a penicillin bottle, and the liquid in the liquid separator sequentially passes through the peristaltic pumps, the filter and the filling needle head and enters the penicillin bottle. Filling was performed at 4 ml/bottle. The filling amount was checked during filling, and the fluctuation range of the filling amount was ±2%.
(4) And (5) plugging the penicillin bottle, capping, checking with eyes, and packaging to obtain a finished product.
(5) Taking 20 bottles of finished products of the preparation, and checking visible foreign matters according to the lamp inspection method of the general rule 0904 visible foreign matter checking method of Chinese pharmacopoeia (2015 edition).
The 20 bottles of the finished product particles are grade A, no visible particles exist, and the requirements of Chinese pharmacopoeia (2015 edition) are met.
In accordance with the present disclosure, while the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention.
The disclosures of all documents cited herein are hereby incorporated by reference to the extent that they provide exemplary, procedural and other details supplementary to those set forth herein.
Claims (10)
1. An apparatus for producing a recombinant protein composition comprising the following elements: a stock solution liquid storage bag or a freeze thawing bag (1) for storing a stock solution of a recombinant protein composition, a front peristaltic pump (2), a front filter (3), a middle liquid storage bag (4), a liquid separator (5), a peristaltic pump (6) for filling and a filling needle (8).
2. The apparatus according to claim 1, wherein: stock solution reservoir bag or freeze thawing bag (1), front end peristaltic pump (2), two front end filters, intermediate stock solution bag (4), knockout (5), peristaltic pump (6) are used in the filling, filling needle (8), each component connects gradually.
3. The apparatus according to any one of claims 1-2, wherein: the dispenser (5) is connected to n peristaltic pumps (6) connected in parallel to each other, each peristaltic pump (6) being connected to a respective filling needle (8), wherein n is an integer from 1 to 24, for example n may be 1,2,3,4,5,6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, n is preferably an integer from 1 to 12, for example n may be 1,2,3,4,5,6,7,8,9, 10, 11, 12, most preferably n=6.
4. A device according to any one of claims 1-3, characterized in that: the device is characterized in that the raw liquid storage bag or the raw liquid freezing and thawing bag (1), the front peristaltic pump (2), the two front filters, the middle liquid storage bag (4), the liquid separator (5), the peristaltic pump (6) for filling and/or other elements can be inserted or added between or at two ends of each two elements of the filling needle (8), and preferably, the rear filter (7) is respectively added after each peristaltic pump for filling which is mutually connected in parallel, so that each peristaltic pump for filling which is mutually connected in parallel is respectively connected with one rear filter and one filling needle in sequence.
5. The apparatus according to any one of claims 1-4, wherein: the front end filter pore size is 0.1 μm to 1 μm, preferably 0.1 μm,0.22 μm,0.45 μm, most preferably 0.22 μm; and/or the pore size of the back-end filter is 0.1 μm to 1 μm, preferably 0.1 μm,0.22 μm,0.45 μm, most preferably 0.22 μm.
6. A method of preparing a recombinant protein composition, the method comprising filling a recombinant protein composition, the filling a recombinant protein composition comprising: i) Filtering the recombinant protein composition using a filter, ii) filling the composition into penicillin bottles using a filling needle.
7. The method according to claim 6, wherein: the filter is a rear-end filter, and is directly connected with the filling needle through a pipeline.
8. The method according to any one of claims 6 to 7, characterized in that the preparation of the recombinant protein composition is carried out using the apparatus according to any one of claims 1 to 5.
9. The method according to any one of claims 6-8, wherein: the recombinant protein is selected from a coagulation factor or an IgG4 antibody, preferably recombinant human coagulation factor VIIa or sodium Wu Liyou monoclonal antibody.
10. Use of the device according to any one of claims 1-5 for the preparation, filling, packaging and/or production of recombinant protein pharmaceutical compositions.
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| JPH066839Y2 (en) * | 1988-04-18 | 1994-02-23 | エーザイ株式会社 | Liquid transport pipe of liquid collector |
| CN2533067Y (en) * | 2000-12-22 | 2003-01-29 | 李力 | Bacterial filling device |
| US20020124526A1 (en) * | 2001-03-12 | 2002-09-12 | Lewis James D. | Albumin in a flexible polymeric container |
| US20090280229A1 (en) * | 2008-05-07 | 2009-11-12 | Constantine Wendy L | Method and apparatus for manufacturing protein beverages |
| CN103863984A (en) * | 2012-12-14 | 2014-06-18 | 山东绿叶制药有限公司 | Sterile filling infusion system with automatic liquid feeding control |
| SG10201709131UA (en) * | 2013-03-08 | 2017-12-28 | Genzyme Corp | Continuous purification of therapeutic proteins |
| CN104510710A (en) * | 2013-09-30 | 2015-04-15 | 瑞阳制药有限公司 | Small-volume pefloxacin mesylate freeze-dried powder injection, preparation method and producing device thereof |
| CN104353282A (en) * | 2014-11-12 | 2015-02-18 | 江苏省原子医学研究所 | Filling closed system sterile-filtration equipment |
| CN107287265B (en) * | 2016-03-31 | 2018-09-14 | 正大天晴药业集团南京顺欣制药有限公司 | A kind of method of Prepare restructuring human blood coagulation factors VIII |
| CN206142204U (en) * | 2016-08-19 | 2017-05-03 | 石狮海星食品有限公司 | Plain and compound oral liquid filling equipment of collagen of king crab |
| CN206511159U (en) * | 2016-12-07 | 2017-09-22 | 河南泰丰生物科技有限公司 | A kind of liquid medicine aseptic filling systems |
| JP2018150289A (en) * | 2017-03-15 | 2018-09-27 | 株式会社日立製作所 | Protein purification methods and protein purification apparatus |
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| CN208021799U (en) * | 2018-03-28 | 2018-10-30 | 郑州国食科技有限公司 | A kind of filling production metering device for wheat germ globulin oral solution |
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