WO2017065559A1 - Procédé de production de protéine de fusion présentant un domaine fc d'igg - Google Patents

Procédé de production de protéine de fusion présentant un domaine fc d'igg Download PDF

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Publication number
WO2017065559A1
WO2017065559A1 PCT/KR2016/011561 KR2016011561W WO2017065559A1 WO 2017065559 A1 WO2017065559 A1 WO 2017065559A1 KR 2016011561 W KR2016011561 W KR 2016011561W WO 2017065559 A1 WO2017065559 A1 WO 2017065559A1
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Prior art keywords
culture
protein
cell
cells
temperature
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PCT/KR2016/011561
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English (en)
Korean (ko)
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박순재
정혜신
김진환
유선아
조정수
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(주)알테오젠
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Priority claimed from KR1020160132633A external-priority patent/KR101936049B1/ko
Priority to EP16855776.7A priority Critical patent/EP3363811B1/fr
Priority to RU2018117705A priority patent/RU2732237C2/ru
Priority to ES16855776T priority patent/ES2945537T3/es
Priority to MX2018004580A priority patent/MX2018004580A/es
Priority to CN201680059726.XA priority patent/CN108137672B/zh
Priority to CA3001977A priority patent/CA3001977C/fr
Priority to PL16855776.7T priority patent/PL3363811T3/pl
Application filed by (주)알테오젠 filed Critical (주)알테오젠
Priority to BR112018007590-6A priority patent/BR112018007590A2/pt
Priority to JP2018519736A priority patent/JP6783305B2/ja
Priority to EP22186197.4A priority patent/EP4098659A1/fr
Priority to US15/767,806 priority patent/US11414476B2/en
Priority to AU2016339642A priority patent/AU2016339642B2/en
Priority to CN202210985550.8A priority patent/CN115247196A/zh
Publication of WO2017065559A1 publication Critical patent/WO2017065559A1/fr
Priority to US17/859,466 priority patent/US20220348636A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/71—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators

Definitions

  • the present invention relates to a fusion protein having a human immunoglobulin G (IgG) Fc domain (particularly, a fusion of extracellular domain of human vascular endothelial growth factor (VEGF) receptor and human immunoglobulin G (IgG) Fc domain).
  • the present invention relates to a method for producing a protein (eg, Aflibercept).
  • VEGF Vascular Endothelial Growth Factor
  • RNA aptamer RNA aptamer
  • Ranibizumab monoclonal IgG antibody fragment (Fab)
  • Bevacizumab monoclonal IgG antibody
  • Aflibercept VEGFR1 and VEGFR2 fused with IgG1 Fc
  • the present invention has been made to solve the above problems, and provides a method for producing an IgG Fc fusion protein for increasing the amount of protein expression.
  • Another object of the present invention is to provide a method for producing a target protein comprising culturing a cell producing the target protein by the above-described production method.
  • It is still another object of the present invention to provide a pharmaceutical composition comprising a therapeutic protein prepared by the above-described preparation method and a pharmaceutically acceptable carrier.
  • the present invention was confirmed that the productivity and quality of the fusion protein is improved by optimizing the culture conditions of the cells producing the fusion protein having the IgG Fc domain. Specifically, after culturing at a normal culture temperature (35.0 °C ⁇ 38.0 °C) for a period of time, the culture temperature is reduced to 28.0 °C ⁇ 35.0 °C by culturing, thereby increasing the productivity of the fusion protein and aggregates (aggregates) of the fusion protein ) Inhibition of production was confirmed and based on this, the present invention was completed.
  • the present invention provides a method for producing a protein in which a soluble extracellular domain of vascular endothelial growth factor (VEGF) receptor and a human immunoglobulin G Fc domain are fused in cell culture.
  • VEGF vascular endothelial growth factor
  • a method of culturing cells at a reduced temperature of less than 28.0 ° C. to less than 35.0 ° C. is provided to increase the expression level of the fusion protein.
  • the fusion protein prepared by the above method may be one in which aggregates are reduced.
  • the cell culture may be a large-scale cell culture.
  • the cell culture is a batch culture (batch culture), repeated batch culture (repeated batch culture), fed-batch culture, repeated fed-batch culture (repeated fed-batch culture) It may be any one selected from the group consisting of, continuous culture (peruous culture) and perfusion culture (perfusion culture).
  • the cell culture may be a fed-batch cell culture.
  • the cell may be a mammalian cell.
  • the mammalian cell may be a CHO cell.
  • the CHO cells may be any one cell line selected from the group consisting of DG44, DXB-11, K-1 and CHO-S.
  • the culture temperature before the temperature change from the start of the culture may include a temperature range of less than 33.0 °C to 38.0 °C.
  • the reduced temperature may be 30.0 °C to 34.0 °C.
  • the incubation period may be from 1 to 5 days before the temperature change from the start of the culture.
  • the incubation period after the temperature decrease may be 2 to 15 days.
  • the sum of the culture period before the temperature change and the culture period after the temperature change may be 3 days or more.
  • the soluble extracellular domain of the VEGF receptor may comprise immunoglobulin-like domain 2 of the first VEGF receptor and immunoglobulin-like domain 3 of the second VEGF receptor.
  • the produced protein may be a therapeutic protein.
  • the present invention also provides a method for producing a target protein comprising culturing a cell producing the target protein by the above-described production method.
  • the cell producing the target protein may further comprise the step of recovering the target protein from the culture medium.
  • the target protein may be a therapeutic protein.
  • the present invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutic protein prepared by the above-described preparation method and a pharmaceutically acceptable carrier.
  • the present invention further comprises the step of culturing cells producing a fusion protein having an IgG Fc domain at a reduced culture temperature, thereby increasing cell growth and cell viability, increasing the expression level of the fusion protein, and aggregates. Inhibiting production, consequently increasing the productivity and improving the quality of the fusion protein, allowing mass production and supply of the fusion protein.
  • Aflibercept 1 is a graph analyzing the change of cell growth and cell viability according to the culture temperature of cells producing Aflibercept (Aflibercept).
  • FIG. 2 shows the integrated viable cell count (Y axis; IVC [normalized 10 9 cells ⁇ day) normalized to IVC of cells producing Aflibercept over time (X axis; incubation time [day]). / L]).
  • FIG 3 is a graph showing a specific production rate change with the culture temperature of the cells producing Aflibercept (Aflibercept).
  • Figure 4 is a graph of the change in the expression amount according to the culture temperature of the cells producing Aflibercept (HPLC) by high performance liquid chromatography.
  • FIG. 5 is a graph analyzing the change of cell growth and cell viability according to the cold culture temperature of the cells producing Aflibercept (Aflibercept).
  • FIG. 6 is a graph of HPLC (high performance liquid chromatography) analysis of changes in expression level according to cold culture temperature of cells producing Aflibercept.
  • FIG. 7 is a graph illustrating changes in cell growth and cell viability according to culture temperature of cells producing Aflibercept in a 2L bioreactor.
  • FIG. 8 shows the integrated viable cell count (Y axis; IVC [normalized to IVC) of cells producing Aflibercept over time (X axis; incubation time [day]) in a 2L bioreactor. Normalized 10 9 cells ⁇ day / L]).
  • FIG. 9 is a graph showing changes in specific production rate according to the culture temperature of cells producing Aflibercept in a 2L bioreactor.
  • FIG. 10 is a graph of protein A-HPLC (High Performance Liquid Chromatography) analyzing the change in the expression level according to the culture temperature of cells producing Aflibercept in a 2L bioreactor.
  • FIG. 11 is a graph of SE-HPLC (size exclusion high performance liquid chromatography) analysis of changes in aggregated protein according to the culture temperature of cells producing Aflibercept in a 2L bioreactor.
  • the present inventors incubated at a normal culture temperature (35.0 ° C. to 38.0 ° C.) for a certain period of time, and then cultured by reducing the culture temperature to 28.0 ° C. to 35.0 ° C., thereby producing a fusion protein having an IgG (Immunoglobulin G) Fc domain. It was confirmed that this increase and the production of aggregates (aggregates) of the fusion protein is suppressed, and the solution of the above-described problem was sought by providing a method for producing a fusion protein having an IgG Fc domain, the protein expression amount is increased in cell culture.
  • a normal culture temperature 35.0 ° C. to 38.0 ° C.
  • the method for producing a fusion protein having an IgG Fc domain of the present invention increases the cell growth and cell viability by performing a step of culturing the cells producing the fusion protein having an IgG Fc domain at a reduced culture temperature, thereby increasing the cell growth and cell viability.
  • fusion protein having an IgG Fc domain means a protein bound to an Fc region which is a constant region of human immunoglobulin G (IgG).
  • protein means a peptide bond of several or more amino acids.
  • the "amino acid polymer” may use human VEGF receptors 1 and 2, and preferably the extracellular domains of VEGF receptors 1 and 2 may be used.
  • Fc region may be used as a constant region of the antibody human IgG1, IgG2, IgG3 and IgG4, preferably the Fc region of IgG1 can be used.
  • the present invention provides a method for producing a protein in which a soluble extracellular domain of a Vascular Endothelial Growth Factor (VEGF) receptor and a human immunoglobulin G Fc domain are fused in cell culture.
  • VEGF Vascular Endothelial Growth Factor
  • a human immunoglobulin G Fc domain are fused in cell culture.
  • the cell culture may be a large-scale cell culture
  • the cell culture method may use a conventionally used cell culture method.
  • the cell culture method is not limited to this, but batch culture, repeated batch culture, fed-batch culture, repeated fed-batch culture , At least one selected from the group consisting of continuous culture and perfusion culture.
  • the "batch culture method” is a culture method in which a small amount of seed culture solution is added to the medium, and the cells are grown without adding a medium or releasing the culture medium during the culture.
  • the “continu culture method” is a culture method in which a medium is continuously added during culturing and also continuously discharged.
  • the continuous culture method also includes perfusion culture. Since the "fed-value cultivation method” is halfway between the batch cultivation method and the continuous culturing method, it is also called a semi-batch culture, and the medium is added continuously or sequentially during the cultivation. It is a culture method in which culture medium is discharged but cells are not leaked. In the present invention, any of the above culturing methods may be used.
  • a fed-batch culture method or a continuous culture method may be used, and particularly preferably, a fed-batch culture method may be used.
  • Cells used for the expression of the Fc fusion protein in the present invention can be used without limitation as long as it is a stable cell line capable of continuously expressing the fusion protein, preferably may be a mammalian cell. More preferably, animal culture cells commonly used, such as CHO cells, HEK cells, COS cells, 3T3 cells, myeloma cells, BHK cells, HeLa cells, Vero cells, are used. Cells are preferred.
  • dhfr-CHO cells Proc. Natl. Acad. Sci. USA, 1980, 77, 4216-4220
  • CHO K-1 cells Proc which are CHO cells lacking the DHFR gene
  • CHO cells DG44 strain, DXB-11 strain, K-1 strain or CHO-S strain is preferable, K-1 strain is especially preferable,
  • the introduction of the vector into a host cell is carried out by the calcium phosphate method, DEAE dextran method. , Electroporation, lipofection, or the like can be carried out.
  • the culture temperature from the culture start date until the temperature change can be used by selecting a culture temperature that is commonly used according to the cell type. have.
  • the temperature range typically used for mammalian cell culture may be less than 33.0 ° C to 38.0 ° C, particularly preferably 37.0 ° C.
  • cells overexpressing the recombinant aplibercept were grown at 37.0 ° C. until the temperature change from the start of the culture to grow the cells.
  • the timing of the temperature change is determined by the expression level of the target protein. Specifically, by performing the experiment shown in Example 3, the optimum temperature change timing can be known, but since the final cell density varies depending on the cells used and the culture conditions, it is generally from 1 ⁇ 10 6 cells / ml to Preferred is about 1 ⁇ 10 8 cells / ml.
  • the present invention is a method for suppressing the increase in productivity per cell and aggregates when culturing CHO cells into which a gene encoding a protein is introduced for the purpose of producing the fusion protein.
  • the culture is carried out at a normal culture temperature until 5 days later, and then the culture temperature is reduced.
  • the period from the temperature change to low temperature until the end of the culture is generally 1 day to 30 days, preferably 2 to 15 days.
  • the sum of the incubation period before the temperature change and the incubation period after the temperature change may be 3 days or more.
  • the method for producing the protein by culturing the cells producing the fusion protein characterized in that after culturing at a normal culture temperature for a certain period of time, the culture is continued at a reduced temperature.
  • the typical culture temperature here is generally 33.0 ° C. to 38.0 ° C., which is suitable for cell proliferation of constant temperature animal derived cells, and 37.0 ° C. is most common.
  • a reduced culture temperature means a temperature range lower than a conventional culture temperature, and an optimal reduced culture temperature is a target. It is determined by the expression level of the protein.
  • the optimal alteration temperature can be known through the same experiment as in Example 2, since the final cell density varies depending on the type of cells used or the culture conditions, the optimal reduced culture temperature is preferably 28.0 ° C to 35.0 ° C, More preferably, it may be 30.0 ° C to 34.0 ° C.
  • the soluble extracellular domain of the VEGF receptor is an immunoglobulin-like domain 2 and a second VEGF receptor of the first VEGF receptor.
  • Immunoglobulin-like domain 3 of Specifically, the protein produced by the production method of the present invention may be a therapeutic protein.
  • the cells overexpressing the recombinant aplybercept is 37.0 ° C. with a culture temperature of 37.0 ° C. from the start of the culture to the temperature change until the cell density reaches 8 ⁇ 10 6 cells / mL in the flask. Incubated for Thereafter, the temperature was lowered to 32.0 ° C. and oil culture was carried out according to the feeding schedule of Table 1.
  • the cells growing at a temperature lower than the normal culture temperature in the flask showed higher viability than the cells continuously cultured at 37.0 ° C. (control), thereby increasing the incubation period. As shown, the expression level of the protein was increased.
  • the cell overexpressing the recombinant aplibercept is 37.0 ° C. until the temperature is changed from the start of the culture until the temperature is changed until the density of the cells in the flask reaches 8 ⁇ 10 6 cells / mL. Incubated for days. Thereafter, the temperature was lowered to 30.0 ° C. or 32.0 ° C. or 34.0 ° C., and then cultured with milk according to the feeding schedule of Table 1.
  • the cell concentration is increased compared to other temperatures in the culture at the culture temperature reduced to 34.0 ° C. in the flask, but as shown in FIG. 6, the expression level of the protein in the culture at the culture temperature reduced to 32.0 ° C. is shown. Most increased.
  • the cell overexpressing the recombinant aplibercept is at a culture temperature of 37.0 ° C. from the start of the culture until the temperature change is 4x10 6 cells / mL or 8x10 6 cells / mL in the bioreactor. Incubate for 1 or 2 days until reaching mL. Thereafter, the temperature was lowered to 32.0 ° C. and oil culture was carried out according to the feeding schedule of Table 1.
  • the pH of the culture medium in the bioreactor is different depending on the cells to be cultured, but may generally be pH 6.8 to 7.6, preferably pH 6.8 to 7.4.
  • dissolved oxygen (DO) of the culture medium in the bioreactor is generally 20% to 60%, preferably 30% to 50%, more preferably 40% is used.
  • the method for producing a fusion protein having an IgG Fc domain of the present invention increases the growth and survival rate of cells producing a fusion protein having an IgG Fc domain through optimization of cell culture conditions, thereby increasing the productivity of the fusion protein.
  • the method for producing a fusion protein having an IgG Fc domain of the present invention improves the method of culturing mammalian cells, thereby increasing the productivity of the fusion protein and inducing the aggregation of protein aggregates that affect the quality of the fusion protein. Production can be inhibited to provide fusion proteins with improved quality.
  • the method of the present invention is characterized by increasing the productivity of the desired protein and inhibiting the generation of aggregate components when the fusion protein is produced by culturing cells producing the desired protein. Therefore, the ligand-binding portion of the anti-VEGF receptor will help to improve the purification process by inhibiting the production and aggregation of Aflibercept, a protein fused to the Fc region of IgG1.
  • the present invention also provides a method for producing a target protein comprising culturing a cell producing the target protein by the above-described production method.
  • the method for producing a target protein of the present invention may further include a step of recovering the target protein from the culture medium in which the cells producing the target protein are cultured.
  • the target protein prepared by the method for preparing the target protein may be a therapeutic protein, and the prepared therapeutic protein may be provided in a pharmaceutical composition together with a pharmaceutically acceptable carrier.
  • the pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant may include excipients, disintegrants, sweeteners, binders, coatings, swelling agents, lubricants, lubricants. Solubilizers, such as an agent or a flavoring agent, can be used.
  • the pharmaceutical composition of the present invention may be preferably formulated into a pharmaceutical composition by containing one or more pharmaceutically acceptable carriers in addition to the active ingredient for administration.
  • Acceptable pharmaceutical carriers in compositions formulated in liquid solutions are sterile and physiologically compatible, including saline, sterile water, Ringer's solution, buffered saline, albumin injectable solutions, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.
  • the pharmaceutical composition of the present invention can be administered to various mammals, including humans, according to the kind of therapeutic protein prepared.
  • parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal , Intranasal, intestinal, topical, sublingual, rectal or intravitreal injection administration.
  • Suitable dosages of the pharmaceutical compositions of the present invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and response to response of the patient, Usually a skilled practitioner can easily determine and prescribe a dosage effective for the desired treatment or prophylaxis.
  • Recombinant Aflibercept was cloned as a fusion protein using an improved vector from which shGH, His tag, and TEV sites were removed from pSGHV0 (GenBank Accession No. AF285183), and a unique signal sequence was used It was allowed to secrete extracellularly.
  • the GS system was introduced as a selection marker for constructing a stable cell line expressing the fusion protein.
  • a mouse glutamine synthetase gene was inserted into a vector.
  • Kozac sequence was further inserted into the signal sequence to increase expression.
  • the clone thus prepared was introduced into the CHO-K1 cell line (ATCC, Cat. CCL-61) to proceed with the selection of methionine sulphoximine (MSX) to secure a stable cell line.
  • MSX methionine sulphoximine
  • one of ordinary skill in the art may appropriately select and apply vectors and cell lines which are commonly used according to a given situation.
  • Cells overexpressing the recombinant Aflibercept prepared in the above preparation was inoculated in two 125 mL cells in a Erlenmeyer flask at the same concentration and conditions to which the plant-derived hydrolyzed protein was added and 37.0. Shake incubation in a C 2 CO 2 incubator. The cells were grown in batch culture, and then fed to a fed-batch culture by lowering the temperature to 32.0 ° C. when the cell concentration was about 8 ⁇ 10 6 cells / mL.
  • the feeding schedule for the experimental conditions is summarized in Table 1 below. The feedstock volume is described as a percentage of the starting volume of culture in the bioreactor.
  • IVC Intelligent viable cell
  • Cells overexpressing the recombinant Aflibercept prepared in the above Preparation Example were inoculated in three 125 mL in an Erlenmeyer flask in a medium to which the plant-derived hydrolyzed protein was added at the same concentration and conditions, and 37.0. Shake incubation in a C 2 CO 2 incubator. The cells were grown in batch culture, and when the cell concentration was about 8 ⁇ 10 6 cells / mL, the cells were fed incubated by lowering the temperature to 30.0 ° C., 32.0 ° C. or 34.0 ° C., respectively. The supply schedule for the experimental conditions was performed as shown in Table 1 of Example 1. Various conditions of the cells were measured as described in Example 1.
  • the above results can be produced in high quality by inhibiting the generation of aggregates of Aflibercept represented by the fusion protein through the production method of the protein fused with the soluble extracellular domain and the human IgG Fc domain of the VEGF receptor of the present invention.
  • the expression level can be increased significantly through the incubation at reduced temperature, thereby increasing the productivity.
  • the fusion protein production method of the present invention is optimized for the production of a protein (eg, aflibercept) fused with a soluble extracellular domain of the VEGF receptor and a human IgG Fc domain.
  • a protein eg, aflibercept
  • Different levels of protein can significantly alter the productivity and quality of the protein. Therefore, if the type of target protein to be produced is changed, a process of resetting and verifying optimized conditions for improving quality and productivity will be required.
  • the method for producing a protein in which the soluble extracellular domain and the human IgG Fc domain of the VEGF receptor provided by the present invention are fused includes adding a step of culturing the cells producing the fusion protein at a reduced culture temperature, thereby increasing cell growth and cell viability.
  • the protein produced by the production method of the present invention is a therapeutic protein, which can provide a pharmaceutical composition in a suitable form according to the therapeutic purpose, and thus has great industrial applicability.

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Abstract

La présente invention concerne un procédé de préparation d'une protéine de fusion présentant un domaine Fc d'IgG et, plus particulièrement, un procédé de préparation d'une protéine de fusion présentant un domaine Fc d'IgG, le procédé comprenant en outre une étape de culture de cellules, qui produisent la protéine de fusion, à une température de culture réduite, ce qui permet d'augmenter la croissance cellulaire et la viabilité cellulaire, de manière à augmenter la productivité en protéines de fusion, et d'inhiber la génération d'agrégats de manière à améliorer la qualité et le rendement de production.
PCT/KR2016/011561 2015-10-15 2016-10-14 Procédé de production de protéine de fusion présentant un domaine fc d'igg WO2017065559A1 (fr)

Priority Applications (14)

Application Number Priority Date Filing Date Title
CN202210985550.8A CN115247196A (zh) 2015-10-15 2016-10-14 用于生产具有igg fc结构域的融合蛋白的方法
PL16855776.7T PL3363811T3 (pl) 2015-10-15 2016-10-14 Sposób wytwarzania białka fuzyjnego z domeną fc igg
ES16855776T ES2945537T3 (es) 2015-10-15 2016-10-14 Método para producir proteína de fusión que tiene un dominio Fc de IgG
MX2018004580A MX2018004580A (es) 2015-10-15 2016-10-14 Metodo para producir proteina de fusion con dominio fc de igg.
BR112018007590-6A BR112018007590A2 (pt) 2015-10-15 2016-10-14 método para produzir proteína de fusão que tem domínio fc de igg
CA3001977A CA3001977C (fr) 2015-10-15 2016-10-14 Procede de production de proteine de fusion presentant un domaine fc d'igg
RU2018117705A RU2732237C2 (ru) 2015-10-15 2016-10-14 Способ получения афлиберцепта
EP16855776.7A EP3363811B1 (fr) 2015-10-15 2016-10-14 Procédé de production de protéine de fusion présentant un domaine fc d'igg
CN201680059726.XA CN108137672B (zh) 2015-10-15 2016-10-14 用于生产具有igg fc结构域的融合蛋白的方法
JP2018519736A JP6783305B2 (ja) 2015-10-15 2016-10-14 ＩｇＧ Ｆｃドメインを有する融合タンパク質の生産方法
EP22186197.4A EP4098659A1 (fr) 2015-10-15 2016-10-14 Procédé de production de protéine de fusion dotée d'un domaine fc igg
US15/767,806 US11414476B2 (en) 2015-10-15 2016-10-14 Method for producing fusion protein having IgG Fc domain
AU2016339642A AU2016339642B2 (en) 2015-10-15 2016-10-14 Method for producing fusion protein having IgG Fc domain
US17/859,466 US20220348636A1 (en) 2015-10-15 2022-07-07 Method for producing fusion protein having igg fc domain

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KR20150144330 2015-10-15
KR10-2015-0144330 2015-10-15
KR1020160132633A KR101936049B1 (ko) 2015-10-15 2016-10-13 IgG Fc 도메인을 가지는 융합 단백질의 생산방법
KR10-2016-0132633 2016-10-13

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US15/767,806 A-371-Of-International US11414476B2 (en) 2015-10-15 2016-10-14 Method for producing fusion protein having IgG Fc domain
US17/859,466 Continuation US20220348636A1 (en) 2015-10-15 2022-07-07 Method for producing fusion protein having igg fc domain

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2020517265A (ja) * 2017-04-21 2020-06-18 ユーハン・コーポレイションＹＵＨＡＮ Ｃｏｒｐｏｒａｔｉｏｎ デュアル機能タンパク質およびその誘導体を生産するための方法

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US11560416B2 (en) 2017-04-21 2023-01-24 Yuhan Corporation Method for producing dual function proteins and its derivatives
JP2023036638A (ja) * 2017-04-21 2023-03-14 ユーハン・コーポレイション デュアル機能タンパク質およびその誘導体を生産するための方法

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