CN1138564C - 流感疫苗 - Google Patents
流感疫苗 Download PDFInfo
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- CN1138564C CN1138564C CNB981080863A CN98108086A CN1138564C CN 1138564 C CN1138564 C CN 1138564C CN B981080863 A CNB981080863 A CN B981080863A CN 98108086 A CN98108086 A CN 98108086A CN 1138564 C CN1138564 C CN 1138564C
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Abstract
本发明涉及一种流感表面抗原疫苗，可由在动物细胞培养物上繁殖的流感病毒制得，其宿主细胞DNA含量等于或小于每剂25pg。本发明还涉及一种从在动物细胞培养物上繁殖的流感病毒制备表面抗原蛋白的方法，包括以下步骤：a.用DNA消化酶处理包含完整病毒的细胞培养液，和b.加入阳离子去污剂，然后分离表面抗原蛋白。
Description
本发明涉及由在动物细胞培养物上繁殖的流感病毒制得的流感表面抗原疫苗，和一种由在动物细胞培养物上繁殖的流感病毒制备表面抗原蛋白的方法。
流感病毒的体长大约125nm，含有一个与核蛋白有关的核糖核酸(RNA)的核心，此核心被一层具有脂质双层结构的病毒被膜围绕。病毒被膜的内层主要由基质蛋白质组成，外层含有大部分来源于宿主的类脂物质。
所谓的“表面蛋白质”，即神经氨酸酶(NA)和血细胞凝集素(HA)，表现为病毒体表的刺突。
大部分市售的失活流感疫苗是所谓的“断裂疫苗”或“亚单位疫苗”。
“断裂疫苗”是通过将整个流感病毒经可溶解浓度的去污剂处理，随后除去去污剂和大部分病毒的脂类物质制得。
抗流感“亚单位疫苗”与“断裂疫苗”不同，不含有全部的病毒蛋白质。“亚单位疫苗”富含在接种时可以诱发所需的中和(因此保护)抗体的病毒的表面蛋白。
大部分市售的流感疫苗由在鸡含胚卵上培育的流感病毒得到。然而一般认为，从卵来源制备的流感病毒作为疫苗有一些不利之处：
1.其制备过程由于卵的(微)生物学性质的不定性很易受破坏。
2.如果遇到突然增加的需求，即在感冒严重流行或大流行的情况下，其制备过程由于不能得到大量合格的鸡蛋而完全缺乏产量的适应性。
3.由此制备的疫苗对于已知对鸡和/或鸡蛋蛋白质过敏的人是禁忌的。
对于这些问题的一种解决办法可以求助于从组织培养物制备流感病毒。此制备方法被认为具有许多优点：
1.组织培育的细胞系可以在完全确定的，不含(微)生物污染物的细胞库体系中得到，因此批次间的一致性大大增加，从而得到高质量的产品。
2.在严重的流行病或传染病威胁的情况下，能得到充足疫苗的机会将增加。
3.所得的流感病毒物质将更适合于给药途径的选择(口服、经鼻给药、吸入)。
4.按WHO的观点，这项技术使每年一度的疫苗组合物的推荐可以被推迟(从二月中旬至三月中旬)，可以增进疫苗与流传菌株的匹配。
然而，流感病毒的组织培养也存在一个重要问题，因为从传代细胞系中得到的遗传物质可能仍存在于疫苗中。
这些问题形成一种危险，如不加以克服，将导致管理部门由于安全原因拒绝允许这些流感疫苗上市的请求。如美国食品药品管理局要求用于人体的生物技术产品每次服用剂量中含有的宿主细胞DNA不能超过100pg。
因此，本发明提供一种制备流感病毒表面抗原的方法，由此制得的疫苗是安全的，每次服用剂量中所含的有害遗传物质不超过可被接受的数量，以符合管理部门提出的要求。无论如何，此方法被认为是理想的和令人惊喜地可以制备每一服用剂量所含宿主DNA量大大低于100pg的流感疫苗。
相应地，本发明涉及一种从在动物细胞培养物上繁殖的流感病毒制得的流感表面抗原疫苗，其每次服用剂量中所含宿主细胞DNA等于或小于25pg。
具体地说，本发明提供一种制备表面抗原蛋白的方法，用于通过在动物细胞培养物上繁殖的流感病毒制备这种低DNA的流感疫苗，它包括以下步骤：
a.用一种DNA消化酶处理含有整个病毒的细胞培养液，和
b.加入阳离子去污剂，随后分离表面抗原蛋白。
本发明的方法适用于制备含多种流感病毒株的疫苗，如人流感病毒、猪流感病毒、马流感病毒、鸟流感病毒。
本发明的动物细胞培养物可以含有初级细胞如鸡胚成纤维细胞(CEF)或传代细胞系如犬肾细胞(Madin Darby Canine KidneyCells(MDCK))、中国仓鼠卵巢细胞(CHO)和Vero细胞。
可以直接在发酵罐中用DNA消化酶处理含完整病毒的发酵液，可以选择在细胞培养或病毒繁殖过程中进行。
适合的DNA消化酶是脱氧核糖核酸酶(如按照EC 3.1.21和EC3.1.22分类的)和核酸酶(按照EC3.1.30和3.1.31分类的)。
本发明所用适合的阳离子去污剂主要包括一种具有以下通式的化合物其中R1、R2和R3是相同或不同的，各自代表烷基或芳基，
或R1和R2，与相连的氮原子一起形成5或6元结构的杂环，同时R3代表烷基或芳基，
或R1、R2和R3与相连的氮原子一起形成5或6元结构的杂环，在氮原子处不饱和，R4代表烷基或芳基，并且X代表阴离子。
阳离子去污剂的例子是十六烷基三甲基铵盐，如溴化十六烷基三甲基铵(C.T.A.B.)，和十四烷基三甲基铵盐。合适的去污剂也可以是lipofectine、lipofectamine、DOTMA。
阳离子去污剂可以补充以非离子去污剂，如吐温。
经去污剂处理后表面抗原蛋白的分离可以包括以下步骤：
1.用离心或超滤等方法从表面抗原蛋白中分离核糖核蛋白微粒(体)，和
2.通过去污剂与一种合适的树脂(如Amberlite XAD-4)的疏水性相互作用和/或通过超滤(透析)的方法，从表面抗原蛋白中除去去污剂。
本发明的方法令人惊喜地得到一种含有极低量的来源于动物细胞的DNA的产物。一次剂量中所含DNA低于25pg，在许多样品中甚至很易达到低于每剂10pg。
表面抗原蛋白可以被加工制成流感疫苗，如通过加入缓冲剂(如磷酸盐缓冲液)和/或与其它流感病毒血清型的抗原混合。
进一步制备疫苗过程中可以选择性地经过表面抗原的浓缩。
实施例1A.病毒繁殖
1.在35℃条件下，将种子病毒和细胞在发酵罐中温育2天，使B/Yamagata型抗原的流感病毒在狗肾(Madin Darby CanineKidney(MDCK))细胞(ATCC CCL34)上繁殖。
2.随后加入稀NaOH将发酵液pH提高到8.0，并加入Benzon核酸酶至最终浓度为每升1000单位(1μg)。
3.在35℃条件下再进行4小时的温育。B.病毒分离
1.发酵液通过一种滤孔为0.5μm的深度滤器(depth filter)过滤除去细胞碎屑。
2.随后以截留分子量为300.000的滤膜超滤，将流感病毒浓缩和纯化。
3.在浓缩物中加入蔗糖至终浓度为30％(w/v)，然后加入甲醛至终浓度为0.015％(w/v)。此混合物在2-8℃搅拌72小时。
4.以磷酸盐缓冲盐水将病毒浓缩物稀释5倍，加到含有AmiconCellufine Sulphate的亲合柱上。以磷酸盐缓冲盐水洗涤除去杂质后，用1.5摩尔浓度氯化钠的磷酸盐缓冲盐水溶液洗脱病毒。洗脱物通过截留分子量为300.000的滤膜超滤而被浓缩并脱盐。C.亚单位分离
1.加入非离子去污剂吐温-80至终浓度为300μg/ml，加入溴化十六烷基三甲基铵至终浓度为750μg/ml。将此混合物在4℃搅拌3小时，然后离心以从表面抗原蛋白中分离核糖核蛋白颗粒。
2.上清液加入Amberlite XAD-4树脂在2-8℃搅拌过夜，除去去污剂。过滤除去Amberlite树脂，滤液随后通过0.22μm滤孔的滤器进行无菌过滤。
在以上步骤中，以有效的试验方法对样品中的宿主细胞DNA含量进行分析，此试验基于用一种32 P标记的犬DNA探针进行的狭线印迹杂交。
各制备步骤后DNA的测定结果列于下表(此表中每剂量流感病毒疫苗中的DNA量表示为每50μg血细胞凝集素所含的微微克数(pg/50μg HA))。
表
实施例2
| 样品 | DNA含量 | 所经步骤 |
| 经过48小时温育的发酵液体 | 19.000.000 | A1 |
| 经核酸酶处理的发酵液 | 370.000 | A2 |
| 预过滤后的液体 | 10.000 | B1 |
| 超滤后的浓缩物 | 4350 | B2 |
| 灭活和经亲合层析后的液体 | 4200 | B4 |
| 吐温/C.T.A.B.处理并离心后的上清液 | ＜25 | C1 |
| 除去去污剂后的滤液 | ＜25 | C2 |
病毒的繁殖、纯化、灭活和裂解如实施例1所述。
在病毒繁殖的后几个小时(步骤A2 and A3)，在繁殖培养基的pH(步骤A1)条件下发酵液经Benzon核酸酶处理，对于除去DNA产生相似的结果。
脱氧核糖核酸酶I，虽然需要较高浓度，在去除宿主细胞DNA方面(步骤A1-A3)同样有效。采用其它核酸内切酶可以得到相似的结果。
实施例3
制备过程如实施例1或实施例2所述，只是改用离心法去除细胞碎屑(步骤B1)。
实施例4
制备过程如实施例1、实施例2或实施例3所述，只是改用离心法浓缩和纯化发酵液中的病毒(步骤B2)，使用连续液流离心机如在磷酸缓冲盐水中采用蔗糖梯度法的电核型离心机(RK型)。
实施例5
制备过程如实施例1至实施例4所述，只是将在C1步骤中加入溴化十六烷基三甲基铵溶液改为加入溴化十六烷基砒啶、溴化十四烷基三甲基铵、氯化苯双胺、(benzetonium chloride)，氯化甲基苯双胺(methylbenzethonium chloride，)氯化十烷双胺或硬脂酰二甲基溴化苄铵，除去宿主细胞DNA的效果相似。
实施例6
制备过程如实施例1所述，只是改为在经过亲合柱层析后加入蔗糖，加入甲醛达到最大浓度0.05％(w/v)，混合物搅拌120小时。
实施例7
实施例1、2、3、4、和6的方法同样成功地适用于从流感病毒株B/Harbin、B/Panama、A/Texas(H1N1)、A/Taiwan(H1N1)、A/Johannesburg(H3N2)和A/Wuhan(H3N2)制备低DNA疫苗。
Claims (8)
1.流感表面抗原疫苗，由在动物细胞培养物上繁殖的流感病毒制得，其宿主细胞DNA含量等于或小于每剂25pg。
2.由在动物细胞培养物上繁殖的流感病毒制备表面抗原蛋白的方法，包括以下步骤：
a.用DNA消化酶处理包含完整病毒的细胞培养液，和
b.加入阳离子去污剂，
然后分离表面抗原蛋白。
3.权利要求2所述方法，其中在细胞培养物中在流感病毒繁殖期进行DNA消化酶处理。
4.权利要求2所述方法，其中阳离子去污剂主要包括具有以下通式的化合物：
其中
R1、R2、R3是相同或不同的，各自代表烷基或芳基，
或R1和R2与相连的氮原子一起形成5或6元结构的杂环，同时R3代表烷基或芳基，
或R1、R2和R3与相连的氮原子一起形成5或6元结构的杂环，在氮原子处不饱和，
R4代表烷基或芳基，
X代表一个阴离子。
5.权利要求2所述方法，其中阳离子去污剂主要包括溴化十六烷基三甲基铵。
6.权利要求2所述方法，其中阳离子去污剂可以补充以非离子去污剂。
7.权利要求2所述方法，其中流感病毒是在动物细胞系上繁殖的。
8.权利要求2所述方法，其中流感病毒是在MDCK细胞上繁殖的。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101365480B (zh) * | 2005-11-01 | 2014-11-05 | 诺华疫苗和诊断有限两合公司 | 经由β-丙内酯处理的残留细胞DNA水平降低的细胞衍生病毒疫苗 |
Families Citing this family (98)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE60036952T2 (de) * | 1999-09-24 | 2008-08-07 | Glaxosmithkline Biologicals S.A. | Influenzavirus-impfstoffzusammensetzung zur nasalen anwendung |
| GB9923176D0 (en) * | 1999-09-30 | 1999-12-01 | Smithkline Beecham Biolog | Novel composition |
| CN100415769C (zh) * | 2002-02-07 | 2008-09-03 | 中国科学院过程工程研究所 | 寡聚或多聚亚基蛋白质分离纯化的方法 |
| ES2323454T5 (es) | 2002-04-30 | 2012-11-14 | Oncolytics Biotech Inc. | Procedimiento mejorado de purificación de virus |
| US20060110406A1 (en) * | 2003-02-25 | 2006-05-25 | Medimmune Vaccines, Inc. | Refrigerator-temperature stable influenza vaccine compositions |
| EP2481819A1 (en) * | 2003-02-25 | 2012-08-01 | MedImmune Vaccines, Inc. | Cross flow filtration in the production of stabilized influenza vaccine compositions |
| US7270990B2 (en) * | 2003-06-20 | 2007-09-18 | Microbix Biosystems, Inc. | Virus production |
| US8592197B2 (en) | 2003-07-11 | 2013-11-26 | Novavax, Inc. | Functional influenza virus-like particles (VLPs) |
| US8080255B2 (en) | 2003-07-11 | 2011-12-20 | Novavax Inc. | Functional influenza virus like particles (VLPs) |
| US8992939B2 (en) | 2003-07-11 | 2015-03-31 | Novavax, Inc. | Highly efficient influenza matrix (M1) proteins |
| US8506967B2 (en) | 2003-07-11 | 2013-08-13 | Novavax, Inc. | Functional influenza virus like particles (VLPs) |
| ATE435906T1 (de) * | 2004-02-23 | 2009-07-15 | Crucell Holland Bv | Verfahren zur reinigung von viren |
| CA2559371C (en) | 2004-03-09 | 2014-07-08 | Chiron Corporation | Influenza virus vaccines |
| PL2578229T3 (pl) | 2004-09-09 | 2014-01-31 | Novartis Ag | Zmniejszenie potencjalnego zagrożenia jatrogennego związanego z antygenami w szczepionce |
| US7901921B2 (en) | 2004-10-22 | 2011-03-08 | Oncolytics Biotech Inc. | Viral purification methods |
| CN101076355A (zh) | 2004-11-03 | 2007-11-21 | 希龙公司 | 流感疫苗 |
| CA2592120A1 (en) | 2004-12-23 | 2006-07-06 | Medimmune Vaccines, Inc. | Non-tumorigenic mdck cell line for propagating viruses |
| US20080181911A1 (en) | 2005-03-23 | 2008-07-31 | Emmanuel Jules Hanon | Use Of An Influenza Virus, An Oil-In-Water Emulsion Adjuvant, To Induce Cd4 T-Cell And/Or Improved B-Memory Cell Response |
| DK1869171T4 (en) * | 2005-04-11 | 2016-01-18 | Crucell Holland Bv | Virus cleaning using ultrafiltration |
| US11707520B2 (en) | 2005-11-03 | 2023-07-25 | Seqirus UK Limited | Adjuvanted vaccines with non-virion antigens prepared from influenza viruses grown in cell culture |
| CN102727885A (zh) | 2005-11-04 | 2012-10-17 | 诺华疫苗和诊断有限公司 | 包含颗粒佐剂和免疫增强剂组合的流感疫苗 |
| CA2628424A1 (en) | 2005-11-04 | 2007-05-10 | Novartis Vaccines And Diagnostics S.R.L. | Adjuvanted influenza vaccines including cytokine-inducing agents |
| AU2006310163B2 (en) | 2005-11-04 | 2011-09-15 | Seqirus UK Limited | Influenza vaccine with reduced amount of oil-in-water emulsion as adjuvant |
| AU2006310336B2 (en) | 2005-11-04 | 2011-02-03 | Seqirus UK Limited | Adjuvanted vaccines with non-virion antigens prepared from influenza viruses grown in cell culture |
| EP1951301A2 (en) * | 2005-11-04 | 2008-08-06 | Novartis Vaccines and Diagnostics S.r.l. | Emulsions with free aqueous-phase surfactant for adjuvanting split influenza vaccines |
| EP2478916B1 (en) * | 2006-01-27 | 2020-04-01 | Seqirus UK Limited | Influenza vaccines containing hemagglutinin and matrix proteins |
| WO2007110776A1 (en) | 2006-03-24 | 2007-10-04 | Novartis Vaccines And Diagnostics Gmbh & Co Kg | Storage of influenza vaccines without refrigeration |
| JP2009532352A (ja) | 2006-03-31 | 2009-09-10 | ダブリュエーアールエフ−ウィスコンシン アラムナイ リサーチ ファウンデーション | ワクチンのための高力価組み換えインフルエンザ・ウィルス |
| PL2043682T3 (pl) | 2006-07-17 | 2014-09-30 | Glaxosmithkline Biologicals Sa | Szczepionka przeciw grypie |
| GB0614460D0 (en) * | 2006-07-20 | 2006-08-30 | Novartis Ag | Vaccines |
| ES2694805T7 (es) | 2006-09-11 | 2021-10-21 | Seqirus Uk Ltd | Fabricación de vacunas contra virus de la gripe sin usar huevos |
| EP2069480B1 (en) | 2006-09-15 | 2014-03-19 | MedImmune, LLC | Mdck cell lines supporting viral growth to high titers and bioreactor process using the same |
| EP2121011B1 (en) | 2006-12-06 | 2014-05-21 | Novartis AG | Vaccines including antigen from four strains of influenza virus |
| CA2679843C (en) * | 2007-03-05 | 2018-10-02 | Om Pharma | Bacterial extract for respiratory disorders and process for its preparation |
| JP2010521460A (ja) * | 2007-03-12 | 2010-06-24 | アンティジェン・エクスプレス・インコーポレーテッド | 癌免疫療法におけるＩｉ−ＲＮＡｉ関与Ｉｉ抑制 |
| US9452209B2 (en) | 2007-04-20 | 2016-09-27 | Glaxosmithkline Biologicals Sa | Influenza vaccine |
| US9474798B2 (en) | 2007-06-18 | 2016-10-25 | Wisconsin Alumni Research Foundation | Influenza M2 protein mutant viruses as live influenza attenuated vaccines |
| CN101784283A (zh) | 2007-06-27 | 2010-07-21 | 诺华有限公司 | 低添加流感疫苗 |
| EP2772267B1 (en) * | 2007-08-27 | 2016-04-27 | Longhorn Vaccines and Diagnostics, LLC | Immunogenic compositions and methods |
| GB0810305D0 (en) | 2008-06-05 | 2008-07-09 | Novartis Ag | Influenza vaccination |
| JP5518041B2 (ja) | 2008-03-18 | 2014-06-11 | ノバルティス アーゲー | インフルエンザウイルスワクチン抗原の調製における改良 |
| WO2010036760A1 (en) | 2008-09-24 | 2010-04-01 | Medimmune, Llc | Methods for cultivating cells, propagating and purifying viruses |
| WO2010052214A2 (en) * | 2008-11-05 | 2010-05-14 | Glaxosmithkline Biologicals S.A. | Novel method |
| CN102438651A (zh) | 2009-02-10 | 2012-05-02 | 诺华有限公司 | 用于大流行相关毒株的流感疫苗方案 |
| EP2396031A1 (en) | 2009-02-10 | 2011-12-21 | Novartis AG | Influenza vaccines with increased amounts of h3 antigen |
| DK2396032T3 (en) | 2009-02-10 | 2016-12-19 | Seqirus Uk Ltd | Influenza vaccines with reduced amounts of squalene |
| WO2010125461A1 (en) | 2009-04-27 | 2010-11-04 | Novartis Ag | Adjuvanted vaccines for protecting against influenza |
| CN102597246B (zh) | 2009-05-21 | 2014-02-26 | 诺华股份有限公司 | 使用非内源pol I启动子的反向遗传 |
| WO2010144797A2 (en) | 2009-06-12 | 2010-12-16 | Vaccine Technologies, Incorporated | Influenza vaccines with enhanced immunogenicity and uses thereof |
| US9907746B2 (en) | 2009-07-06 | 2018-03-06 | Variation Biotechnologies, Inc. | Methods for preparing vesicles and formulations produced therefrom |
| EP2451950B9 (en) | 2009-07-06 | 2016-11-23 | Variation Biotechnologies Inc. | Methods for preparing vesicles and formulations produced therefrom |
| EP2459722B1 (en) | 2009-07-31 | 2017-09-06 | Seqirus UK Limited | Reverse genetics systems |
| SG178904A1 (en) | 2009-09-10 | 2012-04-27 | Novartis Ag | Combination vaccines against respiratory tract diseases |
| WO2011048560A1 (en) | 2009-10-20 | 2011-04-28 | Novartis Ag | Improved reverse genetics methods for virus rescue |
| EP2493912B1 (en) | 2009-10-26 | 2020-07-29 | Wisconsin Alumni Research Foundation | High titer recombinant influenza viruses with enhanced replication in vero cells |
| US10130697B2 (en) | 2010-03-23 | 2018-11-20 | Wisconsin Alumni Research Foundation (Warf) | Vaccines comprising mutant attenuated influenza viruses |
| CN103025350A (zh) | 2010-05-21 | 2013-04-03 | 诺华有限公司 | 流感病毒的重配方法 |
| EA201291404A1 (ru) | 2010-06-01 | 2013-05-30 | Новартис Аг | Концентрация и лиофилизация вакцинных антигенов гриппа |
| EA201291437A1 (ru) | 2010-06-01 | 2013-05-30 | Новартис Аг | Концентрация вакцинных антигенов без лиофилизации |
| CN103096922B (zh) | 2010-07-06 | 2019-08-06 | 变异生物技术公司 | 用于治疗流行性感冒的组合物和方法 |
| WO2012023044A1 (en) | 2010-08-20 | 2012-02-23 | Novartis Ag | Soluble needle arrays for delivery of influenza vaccines |
| CN103501811B (zh) | 2011-01-13 | 2016-03-30 | 变异生物技术公司 | 用于治疗病毒感染的组合物和方法 |
| CA2862871C (en) | 2011-01-13 | 2020-09-22 | Variation Biotechnologies Inc. | Methods for preparing vesicles and formulations produced therefrom |
| EP2768528A1 (en) | 2011-10-20 | 2014-08-27 | Novartis AG | Adjuvanted influenza b virus vaccines for pediatric priming |
| CA2890084C (en) | 2011-11-18 | 2021-05-04 | Maura Ellen Campbell | Synthetic derivatives of mpl and uses thereof |
| CN104302323A (zh) | 2012-01-12 | 2015-01-21 | 变异生物技术公司 | 用于治疗病毒感染的组合物和方法 |
| EP4008354A1 (en) | 2012-01-27 | 2022-06-08 | Variation Biotechnologies Inc. | Methods and compositions for therapeutic agents |
| AU2013205478B9 (en) | 2012-03-02 | 2014-11-20 | Seqirus UK Limited | Influenza virus reassortment |
| JP2015526062A (ja) | 2012-06-04 | 2015-09-10 | ノバルティス アーゲー | 改善された安全性試験 |
| GB201218195D0 (en) | 2012-10-10 | 2012-11-21 | Istituto Zooprofilattico Sperimentale Delle Venezie | Composition |
| MX2015006927A (es) | 2012-12-03 | 2016-02-05 | Novartis Ag | Redistribucion de virus de la influenza. |
| US10232031B2 (en) | 2013-03-13 | 2019-03-19 | Seqirus UK Limited | Influenza virus reassortment |
| US20140335116A1 (en) | 2013-05-10 | 2014-11-13 | Novartis Ag | Avoiding narcolepsy risk in influenza vaccines |
| DE202013005130U1 (de) | 2013-06-05 | 2013-09-10 | Novartis Ag | Influenza Virus Reassortierung |
| DE202013005100U1 (de) | 2013-06-05 | 2013-08-26 | Novartis Ag | Influenza Virus Reassortierung |
| MX2015016755A (es) | 2013-06-06 | 2016-04-13 | Novartis Ag | Reagrupamiento del virus de influenza. |
| US9950057B2 (en) | 2013-07-15 | 2018-04-24 | Wisconsin Alumni Research Foundation (Warf) | High titer recombinant influenza viruses with enhanced replication in MDCK or vero cells or eggs |
| RU2535153C1 (ru) * | 2013-09-04 | 2014-12-10 | Федеральное государственное унитарное предприятие "Санкт-Петербургский научно-исследовательский институт вакцин и сывороток и предприятие по производству бактерийных препаратов" Федерального медико-биологического агентства России (ФГУП СПбНИИВС ФМБА России) | Способ получения высокоочищенных вирионных концентратов |
| CA2930634C (en) * | 2013-11-15 | 2023-05-09 | Novartis Ag | Removal of influenza nuclear protein (np) from influenza virus preparations |
| WO2015196150A2 (en) | 2014-06-20 | 2015-12-23 | Wisconsin Alumni Research Foundation (Warf) | Mutations that confer genetic stability to additional genes in influenza viruses |
| EP2966093A1 (en) | 2014-07-07 | 2016-01-13 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Process for the preparation of magnetic sulfated cellulose particles, magnetic sulfated cellulose particles and their use |
| US10633422B2 (en) | 2015-06-01 | 2020-04-28 | Wisconsin Alumni Research Foundation (Warf) | Influenza virus replication by inhibiting microRNA lec7C binding to influenza viral cRNA and mRNA |
| RU2614127C2 (ru) * | 2015-06-04 | 2017-03-22 | Общество с ограниченной ответственностью "НТфарма" | Способ получения концентрата рекомбинантных псевдоаденовирусных частиц, экспрессирующих ген гемагглютинина вируса гриппа A/California/07/2009(H1N1) |
| JP2018524323A (ja) | 2015-06-26 | 2018-08-30 | セキラス ユーケー リミテッド | 抗原がマッチしたインフルエンザワクチン |
| WO2017007839A1 (en) | 2015-07-06 | 2017-01-12 | Wisconsin Alumni Research Foundation (Warf) | Improved influenza virus replication for vaccine development |
| JP6830070B2 (ja) | 2015-07-07 | 2021-02-17 | セキラス ユーケー リミテッドＳｅｑｉｒｕｓ ＵＫ Ｌｉｍｉｔｅｄ | インフルエンザ効力アッセイ |
| AU2017221444B2 (en) | 2016-02-19 | 2021-07-29 | Wisconsin Alumni Research Foundation (Warf) | Improved influenza B virus replication for vaccine development |
| EP3601543A4 (en) | 2017-03-30 | 2021-01-27 | Merck Sharp & Dohme Corp. | ADDING NUCLEASES DIRECTLY TO CELL CULTURE TO FACILITATE DIGESTION AND RELEASE OF HOST CELL NUCLEIC ACIDS |
| RU2670001C1 (ru) * | 2017-12-25 | 2018-10-17 | Федеральное государственное бюджетное учреждение науки Институт биохимии и физиологии растений и микроорганизмов Российской академии наук | Способ получения белков клеточной поверхности |
| EP3737750A4 (en) | 2018-01-09 | 2021-11-17 | Synthetic Biologics, Inc. | ALKALINE PHOSPHATASE AGENTS FOR THE TREATMENT OF NEURODevelopmental Disorders |
| US11654184B2 (en) | 2018-03-20 | 2023-05-23 | Theriva Biologics, Inc. | Alkaline phosphatase agents for treatment of radiation disorders |
| US11638699B2 (en) | 2018-03-20 | 2023-05-02 | Theriva Biologics, Inc. | Intestinal alkaline phosphatase formulations |
| EP3914295A2 (en) | 2019-01-23 | 2021-12-01 | Yoshihiro Kawaoka | Mutations that confer genetic stability to additional genes in influenza viruses |
| EP3921413A1 (en) | 2019-02-08 | 2021-12-15 | Wisconsin Alumni Research Foundation (WARF) | Humanized cell line |
| US11390649B2 (en) | 2019-05-01 | 2022-07-19 | Wisconsin Alumni Research Foundation (Warf) | Influenza virus replication for vaccine development |
| JP2022551805A (ja) | 2019-08-27 | 2022-12-14 | ウィスコンシン アルムニ リサーチ ファンデイション | 卵内複製のための安定化されたｈａを持つ組換えインフルエンザウイルス |
| CN115380107A (zh) | 2019-11-18 | 2022-11-22 | 思齐乐私人有限公司 | 用于产生重配流感病毒的方法 |
| KR102444684B1 (ko) * | 2020-04-29 | 2022-09-16 | 에스케이바이오사이언스(주) | 일회용 배양 공정 시스템을 이용한 인플루엔자 바이러스 생산 방법 |
Family Cites Families (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CH589453A5 (zh) * | 1974-01-14 | 1977-07-15 | Sandoz Ag | |
| US4500513A (en) * | 1979-05-15 | 1985-02-19 | Miles Laboratories, Inc. | Influenza vaccine production in liquid cell culture |
| US4317811A (en) * | 1980-09-11 | 1982-03-02 | Merck & Co., Inc. | Herpes simplex type 1 subunit vaccine |
| DE3237313A1 (de) * | 1982-10-08 | 1984-04-12 | Werner Heese | Verfahren zur gewinnung und reinigung antigenhaltiger loesungen, insbesondere fuer influenza-viren, aus antigenhaltiger allantoisfluessigkeit |
| US4783411A (en) * | 1984-10-22 | 1988-11-08 | Janis Gabliks | Influenza-A virus vaccine from fish cell cultures |
| US5173418A (en) * | 1985-05-10 | 1992-12-22 | Benzon Pharma, A/S | Production in Escherichia coli of extracellular Serratia spp. hydrolases |
| US5006472A (en) * | 1988-06-03 | 1991-04-09 | Miles Inc. | Enzymatic purification process |
| KR0148119B1 (ko) * | 1989-02-07 | 1998-08-17 | 심 파쓰 | B형 간염백신의 제조방법 및 정제법 |
| ATE132160T1 (de) * | 1989-03-29 | 1996-01-15 | Univ New York State Res Found | Verfahren zur reinigung von aussenmembran-protein von haemophilus influenza |
| DZ1706A1 (fr) * | 1992-08-07 | 2002-02-17 | Merck & Co Inc | Vaccin contre le virus de l'hepatite a. |
| HRP950097A2 (en) * | 1994-03-08 | 1997-06-30 | Merck & Co Inc | Hepatitis a virus culture process |
| DK1213030T3 (da) * | 1994-11-10 | 2009-07-20 | Baxter Healthcare Sa | Fremgangsmåde til fremstilling af biologiske produkter i protein-fri kultur |
| US5681746A (en) * | 1994-12-30 | 1997-10-28 | Chiron Viagene, Inc. | Retroviral delivery of full length factor VIII |
| CA2228128C (fr) * | 1995-08-01 | 2011-03-29 | Pasteur Merieux Serums & Vaccins | Procede de production industrielle d'un vaccin contre l'encephalite japonaise et vaccin obtenu |
| JPH11512724A (ja) * | 1995-09-28 | 1999-11-02 | ユニバーシティ オブ ピッツバーグ オブ ザ コモンウェルス システム オブ ハイヤー エデュケーション | 標的性の粒子性遺伝子を用いた免疫処置による細胞治療の免疫反応の促進 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101365480B (zh) * | 2005-11-01 | 2014-11-05 | 诺华疫苗和诊断有限两合公司 | 经由β-丙内酯处理的残留细胞DNA水平降低的细胞衍生病毒疫苗 |
| CN104474543A (zh) * | 2005-11-01 | 2015-04-01 | 诺华疫苗和诊断有限两合公司 | 经由β-丙内酯处理的残留细胞DNA水平降低的细胞衍生病毒疫苗 |
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