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1. 发明申请

WO2021047804A1 IN VITRO METHOD FOR TRANSDUCTION OF T CELLS IN THE PRESENCE OF MALIGNANT CELLS 审中-公开
公开(公告)号：WO2021047804A1
公开(公告)日：2021-03-18
申请号：PCT/EP2020/069484
申请日：2020-07-10
申请人： MILTENYI BIOTEC B.V. & CO. KG
发明人： CORDES, Nicole , SCHASER, Thomas , KAISER, Andrew
IPC分类号： A61K39/00 , A61K35/17 , C07K16/44 , C07K16/28 , C07K16/00 , C07K14/725 , A61K39/395 , A61P35/00
摘要： The present invention provides an in-vitro method of reducing the efficiency of transducing malignant cells of the blood system of a subject that are not derived from T cells with lentiviral vector particles without reducing the efficiency of transducing T cells in a sample comprising T cells and said malignant cells. A combination of compositions comprising a first composition and a second composition is also disclosed, wherein said first composition comprises i) transduced T cells of a subject, wherein said transduced T cells express a CAR comprising an antigen binding domain, wherein the antigen binding domain of said CAR binds specifically to a tag of a tagged polypeptide, and ii) non-transduced malignant cells of the blood system of said subject, and wherein said second composition comprises said tagged polypeptide, wherein said tagged polypeptide binds specifically to an antigen expressed on the surface of said malignant cells. Alternatively, the transduced T cells of said first composition may comprise a nucleic acid encoding a CAR and an inducible gene expression system, and said second composition may comprise an induction agent inducing said gene system.

2. 发明申请

WO2020152197A1 A COMBINATION OF COMPOSITIONS FOR ELIMINATION AND ENHANCED ENGRAFTMENT OF HEMATOPOIETIC STEM CELLS IN THE BONE MARROW OF A SUBJECT 审中-公开
公开(公告)号：WO2020152197A1
公开(公告)日：2020-07-30
申请号：PCT/EP2020/051462
申请日：2020-01-22
申请人： MILTENYI BIOTEC B.V. & CO. KG
发明人： PAPANIKOLAOU, Eleni , MILTENYI, Stefan , BOSIO, Andreas , ASSENMACHER, Mario , KAISER, Andrew
IPC分类号： A61K35/17 , A61K35/28 , A61P35/00
摘要： The present invention provides a combination of compositions comprising i) a composition comprising I) a population of T cells, NK cells or cytotoxic immune effector cells comprising a chimeric antigen receptor specific for a stem cell antigen and/or, II) α) a population of T cells, NK cells or cytotoxic immune effector cells comprising a chimeric antigen receptor specific for a tag of a tagged polypeptide, wherein said tagged polypeptide binds specifically to a stem cell antigen, and β) said tagged polypeptide, and ii) a composition comprising a) a population of CD34+ hematopoietic stem cells, and b) one or more accessory or contributory cell populations selected from the group consisting of myeloid cell lineages expressing CD14, CD11b, CD11c, CD123, CD33; CD36; CD47, CD66b, CD235a, CD146 and CD326. A method applying these compositions to a subject in need thereof are also provided.

3. 发明申请

WO2020025468A1 NON-SPECIFIC NUCLEASES FROM THE GENUS PSEUDOMONAS 审中-公开
公开(公告)号：WO2020025468A1
公开(公告)日：2020-02-06
申请号：PCT/EP2019/070148
申请日：2019-07-26
申请人： MILTENYI BIOTEC B.V. & CO. KG
发明人： ELLEUCHE, Skander , SCHMITZ, Sarah , MILTENYI, Stefan , NÖLLE, Volker
IPC分类号： C12N9/22 , C07K14/21
摘要： The present invention provides a composition comprising an isolated polypeptide having non-specific nuclease (NSN) activity for degrading nucleic acids comprising or consisting of at least 70% amino acid sequence identity to SEQ ID NO:2, wherein said polypeptide has NSN activity in a solution of about 4°C, and a cation complexing agent such as EDTA. Said composition may have preferentially a neutral to basic pH. A kit comprising said composition and a method using said composition are also disclosed.

4. 发明申请

WO2017037083A1 METHOD FOR NATURAL KILLER CELL EXPANSION 审中-公开
标题翻译：自然杀伤细胞扩增方法
公开(公告)号：WO2017037083A1
公开(公告)日：2017-03-09
申请号：PCT/EP2016/070455
申请日：2016-08-31
申请人： MILTENYI BIOTEC GMBH
发明人： GRANZIN, Markus , HUPPERT, Volker
IPC分类号： C12N5/0783
CPC分类号： C12N5/0646 , C12N2501/2302 , C12N2501/2315 , C12N2501/2321
摘要： The present invention provides a method for in-vitro culturing and expanding natural killer (NK) cells in a cell culture medium comprising a population of NK cells, the method comprising a) adding an effective concentration of interleukin-21 (IL-21) at the beginning of the culturing process to said medium, b) adding repeatedly an effective concentration of interleukin-2 (IL-2) and/or interleukin-15 (IL-15) to said medium, and c) adding repeatedly feeder cells or membrane particles thereof to said medium, wherein said feeder cells are B cell derived which are EBV immortalized; and wherein said expansion of NK cells in said cell culture medium is maintained for at least 3 weeks.
摘要翻译：本发明提供了在包含NK细胞群的细胞培养基中体外培养和扩增天然杀伤（NK）细胞的方法，所述方法包括：a）将有效浓度的白细胞介素-21（IL-21）加入到培养过程开始于所述培养基，b）向所述培养基反复加入白细胞介素-2（IL-2）和/或白细胞介素-15（IL-15）的有效浓度，和c）重复加入饲养细胞或膜其中所述饲养细胞是B细胞衍生的，其是永生化的EBV; 并且其中所述细胞培养基中NK细胞的所述扩增保持至少3周。

5. 发明申请

WO2023062113A1 METHOD FOR THE GENERATION OF GENETICALLY MODIFIED NK CELLS 审中-公开
公开(公告)号：WO2023062113A1
公开(公告)日：2023-04-20
申请号：PCT/EP2022/078470
申请日：2022-10-13
申请人： MILTENYI BIOTEC B.V. & CO. KG
发明人： ZHANG, Congcong , QUADFLIEG, Melissa , MÖKER, Nina
IPC分类号： C12N5/0783 , C12N15/85
摘要： The present invention provides an in-vitro method for the generation of a population of genetically modified natural killer (NK) cells comprising the steps in the following order: a) obtaining a sample comprising NK cells and other cells, b) enrichment of NK cells from said sample, c) introducing a genetic modifier 1 into said NK cells by electroporation, d) introducing a genetic modifier 2 into said NK cells by transduction, e) expanding said genetically modified NK cells, thereby generating a population of genetically modified NK cells.

6. 发明申请

WO2023057285A1 METHOD FOR TARGETED GENE INSERTION INTO IMMUNE CELLS 审中-公开
公开(公告)号：WO2023057285A1
公开(公告)日：2023-04-13
申请号：PCT/EP2022/077024
申请日：2022-09-28
申请人： MILTENYI BIOTEC B.V. & CO. KG
发明人： LOCK, Dominik , BRANDES, Caroline , ASSENMACHER, Mario , KAISER, Andrew , WILD, Stefan
IPC分类号： C07K14/725 , A61K35/17 , A61K39/00 , C12N9/22 , C12N5/0783 , C12N15/11 , C12N15/90 , C12N15/10 , A61K39/0011 , C07K14/7051 , C12N15/102 , C12N15/907 , C12N2310/20 , C12N2501/2307 , C12N2501/2315 , C12N2510/00 , C12N5/0636
摘要： The present invention provides a method for generating a composition of immune cells expressing a plurality of transgenes under the control of an endogenous promoter of said immune cells, wherein each single immune cell of said composition that underwent the process of allelic exclusion with regard to said endogenous locus expresses only one transgene wherein the transgene encodes a therapeutic protein or therapeutic nucleic acid, thereby generating a plurality of immune cells within said composition of immune cells expressing a plurality of transgenes.

7. 发明申请

WO2019121945A1 METHOD FOR NK CELL TRANSDUCTION 审中-公开
公开(公告)号：WO2019121945A1
公开(公告)日：2019-06-27
申请号：PCT/EP2018/085890
申请日：2018-12-19
申请人： MILTENYI BIOTEC GMBH
发明人： BARI, Rafijul , GRANZIN, Markus , LEUNG, Wing , MÖKER, Nina , HUPPERT, Volker
IPC分类号： C12N15/867
摘要： The present invention discloses an in-vitro method for transferring biological material into activated NK cells with a pseudotyped retroviral vector particle or a virus-like particle thereof, comprising the steps a) activation of NK cells, and b) addition of said pseudotyped retroviral vector particle or virus-like particle thereof to said activated NK cells, wherein said pseudotyped retroviral vector particle or virus-like particle thereof comprises a modified baboon endogenous retrovirus (BaEV) envelope glycoprotein that is able of binding to and fusing with a hematopoietic cell membrane, thereby transferring biological material into said activated NK cells. Preferentially, the activating of NK cells is performed by the addition of a IL-1 family cytokine to the NK cells.

8. 发明申请

WO2018078066A1 ADAPTER CHIMERIC ANTIGEN RECEPTOR EXPRESSING CELLS FOR TARGETING OF MULTIPLE ANTIGENS 审中-公开
标题翻译：表达用于靶向多种抗原的细胞表达嵌合抗体的嵌合抗原受体
公开(公告)号：WO2018078066A1
公开(公告)日：2018-05-03
申请号：PCT/EP2017/077545
申请日：2017-10-27
申请人： MILTENYI BIOTEC GMBH
发明人： KAISER, Andrew , MITTELSTAET, Joerg , HUPPERT, Volker , MILTENYI, Stefan , SCHLEGEL, Patrick , SEITZ, Christian , LANG, Peter , HANDGRETINGER, Rupert
IPC分类号： C07K14/725 , A61K31/4188 , C07K16/28
摘要： The present invention provides an extracellular anti-linker/label epitope chimeric antigen receptor (anti-LLE CAR) comprising I) a linker/label epitope (LLE) binding domain, II) a transmembrane domain, and III) a signal transduction domain, wherein said extracellular LLE binding domain binds a target cell binding molecule (TCBM) comprising i) an antigen binding moiety (ABM), wherein said ABM binds specifically to an antigen expressed on a target cell, ii) a label moiety (LaM), wherein said LaM is a naturally occurring molecule in a subject or a derivative thereof, iii) a linker moiety (LiM) conjugating said ABM and said LaM, thereby forming a linker/label epitope (LLE), wherein said extracellular LLE binding domain binds said LLE with a higher preference than said naturally occurring molecule.
摘要翻译：本发明提供了包含I）接头/标记表位（LLE）结合结构域，II）跨膜结构域和（ii）跨膜结构域的细胞外抗接头/标记表位嵌合抗原受体（抗LLE CAR） III）信号转导结构域，其中所述细胞外LLE结合结构域结合靶细胞结合分子（TCBM），所述靶细胞结合分子包含i）抗原结合部分（ABM），其中所述ABM特异性结合靶细胞上表达的抗原，ii）标记部分（LaM），其中所述LaM是受试者或其衍生物中的天然存在的分子，iii）缀合所述ABM和所述LaM的接头部分（LiM），由此形成接头/标记表位（LLE），其中所述细胞外LLE结合结构域以比所述天然存在的分子更高的优选结合所述LLE。

9. 发明申请

WO2020239866A1 METHOD FOR GENERATION OF GENETICALLY MODIFIED T CELLS 审中-公开
公开(公告)号：WO2020239866A1
公开(公告)日：2020-12-03
申请号：PCT/EP2020/064755
申请日：2020-05-27
申请人： MILTENYI BIOTEC B.V. & CO. KG
发明人： SCHASER, Thomas , KAISER, Andrew , ASSENMACHER, Mario
IPC分类号： C12N5/0783
摘要： The present invention provides a method for the generation of genetically modified T cells comprising the steps a) a sample provided comprising T cells, b) preparation of said sample by centrifugation, c) enrichment of the T cells, d) activation of the T cells using modulatory agents, e) genetic modification of the T cells by transduction with lentiviral vector particles, f) removal of said modulatory agents, thereby generating a sample of genetically modified T cells, wherein said method is performed in equal or less than 144 hours, less than 120 hours, less than 96 hours, less than 72 hours, less than 48 hours, or less than 24 hours. In one embodiment of the invention said enrichment of T cells is performed by magnetic cell separation using magnetic particles that are directly or indirectly coupled to antibodies or antigen binding fragments thereof specific for CD4 and/or CD8 wherein said magnetic particles can be removed from the cells after separation.

10. 发明申请

WO2020169551A1 CELL CULTURE MEDIUM AND METHOD FOR GENERATION OF EPITHELIAL ORGANOIDS FROM EPITHELIAL STEM CELLS 审中-公开
公开(公告)号：WO2020169551A1
公开(公告)日：2020-08-27
申请号：PCT/EP2020/054150
申请日：2020-02-18
申请人： MILTENYI BIOTEC B.V. & CO. KG
发明人： AGORKU, David, Joel , HARDT, Olaf, Thorsten , BOSIO, Andreas , BECKER, Kristin , ECKARDT, Dominik
IPC分类号： C12N5/071
摘要： The present invention provides a cell culture for obtaining an epithelial organoid, the cell culture comprising i) epithelial stem cells, or tissue fragments comprising said epithelial stem cells, ii) a basal medium for animal or human cells, iii) a Bone Morphogenetic Protein (BMP) inhibitor, iv) a mitogenic growth factor selected from the group consisting of epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and keratinocyte growth factor (KGF), v) at least one soluble culture enhancer, wherein said at least one culture enhancer induces correct polarization of the cells in said cell culture within the developing organoid such as a laminin/entactin complex or entactin, and vi) a Wnt agonist if said epithelial stem cells, or tissue fragments comprising said epithelial stem cells are healthy cells, wherein said at least one soluble culture enhancer in said cell culture is a laminin/entactin complex in a concentration between 0.2 mg/ml and 3.4 mg/ml. A cell culture medium, an in-vitro method for obtaining an epithelial organoid, and an epithelial organoid obtained by said method are also disclosed.

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