Method for automated generation of genetically modified T cells

    公开(公告)号:US10131876B2

    公开(公告)日:2018-11-20

    申请号:US15305597

    申请日:2015-04-23

    IPC分类号: C12N5/0783 C12N5/00

    摘要: The present invention provides a process for generation of genetically modified T cells, T cell subsets and/or T cell progenitors comprising the steps: a) providing a cell sample comprising T cells, T cell subsets and/or T cell progenitors b) preparation of the cell sample by centrifugation c) magnetic separation of the T cells, T cell subsets and/or T cell progenitors d) activation of the enriched T cells, T cell subsets and/or T cell progenitors using modulatory agents e) genetic modification of the T cells, T cell subsets and/or T cell progenitors f) expansion of the genetically modified T cells, T cell subsets and/or T cell progenitors in a cultivation chamber g) washing of the cultured T cells, T cell subsets and/or T cell progenitors characterized in that all steps are performed in a closed and sterile cell culture system.

    Indirect antigen-specific T cell recognition assay

    公开(公告)号:US09638689B2

    公开(公告)日:2017-05-02

    申请号:US14201519

    申请日:2014-03-07

    摘要: The present invention provides a method for analyzing simultaneously multiple human antigen-specific cell populations of a sample, the sample comprising B cells and antigen-specific cells, the method comprising a) separation of B cells from said sample, b) dividing the B cells into n sub-samples, c) differentially labeling the B cells of said sub-samples, wherein at least n-1sub-samples are labeled, d) pulsing of the B cells of each sub-sample with single or multiple peptides, e) pooling of the labeled and peptide-pulsed B cells with cells of said sample comprising said antigen-specific cells, f) co-cultivation of the cells of step e), g) flow cytometry analysis of the B cells with regard to their cell number and CD83 expression, thereby determining the potency of said antigen-specific cells in said sample.

    METHOD FOR AUTOMATED GENERATION OF GENETICALLY MODIFIED T CELLS
    6.
    发明申请
    METHOD FOR AUTOMATED GENERATION OF GENETICALLY MODIFIED T CELLS 审中-公开
    自动生成遗传修饰T细胞的方法

    公开(公告)号:US20170037370A1

    公开(公告)日:2017-02-09

    申请号:US15305597

    申请日:2015-04-23

    IPC分类号: C12N5/0783

    摘要: The present invention provides a process for generation of genetically modified T cells, T cell subsets and/or T cell progenitors comprising the steps: a) providing a cell sample comprising T cells, T cell subsets and/or T cell progenitors b) preparation of the cell sample by centrifugation c) magnetic separation of the T cells, T cell subsets and/or T cell progenitors d) activation of the enriched T cells, T cell subsets and/or T cell progenitors using modulatory agents e) genetic modification of the T cells, T cell subsets and/or T cell progenitors f) expansion of the genetically modified T cells, T cell subsets and/or T cell progenitors in a cultivation chamber g) washing of the cultured T cells, T cell subsets and/or T cell progenitors characterized in that all steps are performed in a closed and sterile cell culture system.

    摘要翻译: 本发明提供了一种用于产生遗传修饰的T细胞,T细胞亚群和/或T细胞祖细胞的方法,其包括以下步骤:a)提供包含T细胞,T细胞亚群和/或T细胞祖细胞的细胞样品b) 通过离心分离细胞样品c)磁性分离T细胞,T细胞亚群和/或T细胞祖细胞d)使用调节剂活化富集的T细胞,T细胞亚群和/或T细胞祖细胞e)遗传修饰 T细胞,T细胞亚群和/或T细胞祖细胞f)培养室中遗传修饰的T细胞,T细胞亚群和/或T细胞祖细胞的扩增g)洗涤培养的T细胞,T细胞亚群和/或 T细胞祖细胞的特征在于所有的步骤都是在封闭的和无菌的细胞培养系统中进行的。

    INDIRECT ANTIGEN-SPECIFIC T CELL RECOGNITION ASSAY
    7.
    发明申请
    INDIRECT ANTIGEN-SPECIFIC T CELL RECOGNITION ASSAY 有权
    间接抗原特异性T细胞识别测定

    公开(公告)号:US20140256587A1

    公开(公告)日:2014-09-11

    申请号:US14201519

    申请日:2014-03-07

    IPC分类号: G01N33/52

    摘要: The present invention provides a method for analyzing simultaneously multiple human antigen-specific cell populations of a sample, the sample comprising B cells and antigen-specific cells, the method comprising a) separation of B cells from said sample, b) dividing the B cells into n sub-samples, c) differentially labeling the B cells of said sub-samples, wherein at least n-1sub-samples are labeled, d) pulsing of the B cells of each sub-sample with single or multiple peptides, e) pooling of the labeled and peptide-pulsed B cells with cells of said sample comprising said antigen-specific cells, f) co-cultivation of the cells of step e), g) flow cytometry analysis of the B cells with regard to their cell number and CD83 expression, thereby determining the potency of said antigen-specific cells in said sample.

    摘要翻译: 本发明提供了一种用于同时分析样品的多个人类抗原特异性细胞群的方法,所述样品包含B细胞和抗原特异性细胞,所述方法包括a)从所述样品中分离B细胞,b)将B细胞 c)差异性标记所述亚样品的B细胞,其中至少n-1个样品被标记,d)用单个或多个肽脉冲每个亚样品的B细胞,e) 将标记和肽脉冲的B细胞与包含所述抗原特异性细胞的所述样品的细胞合并,f)步骤e)的细胞的共培养,g)B细胞关于它们的细胞数目的流式细胞术分析 和CD83表达,从而确定所述样品中所述抗原特异性细胞的效力。

    Reversible cell labelling with conjugates having two releasable binding sites

    公开(公告)号:US10890580B2

    公开(公告)日:2021-01-12

    申请号:US15834114

    申请日:2017-12-07

    摘要: The invention is directed to a Method for detecting a target moiety in a sample of biological specimens by: a) providing at least one conjugate with the general formula (I) An-P-Bm-Cq-Xo  (I)  with A: antigen recognizing moiety; P: enzymatically degradable spacer; B: first binding moiety C second binding moiety X: detection moiety; n, m, q, o integers between 1 and 100, wherein B and C are non-covalently bound to each other and A and B are covalently bound to P b) labelling the target moiety recognized by the antigen recognizing moiety A with at least one conjugate c) detecting the labelled target moiety via detecting moiety X d) cleaving Cq-Xo by disrupting the non-covalent bond between Bm and Cq from the labelled target moiety e) cleaving the binding moiety Bm from the labelled target moiety by enzymatically degrading spacer P. The method is useful to identify target moieties on the biological specimens. The biological specimens detected by the conjugate can be subsequently removed from the sample.

    METHOD FOR POLYCLONAL STIMULATION OF T CELLS BY MOBILE NANOMATRICES
    10.
    发明申请
    METHOD FOR POLYCLONAL STIMULATION OF T CELLS BY MOBILE NANOMATRICES 审中-公开
    通过移动纳米粒子对T细胞进行多环刺激的方法

    公开(公告)号:US20150240204A1

    公开(公告)日:2015-08-27

    申请号:US14430875

    申请日:2013-09-25

    IPC分类号: C12N5/0783 A61K35/17

    摘要: The present invention provides a method for polyclonal stimulation of T cells, the method comprising contacting a population of T cells with a nanomatrix, the nanomatrix comprising a) a matrix of mobile polymer chains, and b) attached to said matrix of mobile polymer chains one or more stimulatory agents which provide activation signals to the T cells; thereby activating and inducing the T cells to proliferate; wherein the nanomatrix is 1 to 500 nm in size. At least one first and one second stimulatory agents are attached to the same or to separate mobile matrices. If the stimulatory agents are attached to separate nanomatrices, fine-tuning of nanomatrices for the stimulation of the T cells is possible. Closed cell culture systems also benefit from this method.

    摘要翻译: 本发明提供了一种用于T细胞多克隆刺激的方法,该方法包括使一组T细胞与纳米基质接触,所述纳米基质包含a)可移动聚合物链的基质,和b)连接到所述移动聚合物链的基质上 或更多的刺激剂,其向T细胞提供激活信号; 从而激活和诱导T细胞增殖; 其中纳米线的尺寸为1〜500nm。 至少一个第一和第二刺激剂连接到相同或分离的移动基质。 如果刺激剂连接到分离的纳米片上,则可以对用于刺激T细胞的纳米片进行微调。 闭孔细胞培养系统也受益于这种方法。