公开(公告)号：US10197561B2

公开(公告)日：2019-02-05

申请号：US14944352

申请日：2015-11-18

申请人： Miltenyi Biotec GmbH

发明人： Christian Dose ,  Jennifer Brieden ,  Stefan Miltenyi ,  Thomas Rockel ,  Veronika Gann

IPC分类号： G01N33/542 ,  G01N33/569 ,  G01N33/53 ,  G01N33/537

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) Xn—P—Ym, with X is an detection moiety, P an enzymatically degradable spacer and Y an antigen recognizing moiety and n, m are integers between 1 and 100 and wherein X and Y are covalently bound to P; b) contacting the sample of biological specimens with at least one conjugate, thereby labeling the target moiety recognized by the antigen recognizing moiety Y; c) detecting the target moiety labeled with the conjugate with the detecting moiety X; and d) enzymatically degrading spacer P, thereby cleaving the detection moiety X from the conjugate. 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.

公开(公告)号：US20160347907A1

公开(公告)日：2016-12-01

申请号：US14723505

申请日：2015-05-28

申请人： Miltenyi Biotec GmbH

发明人： Christian Dose

IPC分类号： C08G65/48

CPC分类号： C08G65/48 ,  A61K49/0041 ,  A61K49/0043 ,  A61K49/0045 ,  A61K49/0054 ,  A61K49/0058 ,  C08G65/33306 ,  C08G65/33396 ,  G01N33/582

摘要： The invention is directed to a fluorescent dye according to the general formula I: with C is a core moiety comprising 20 to 200 atoms; S same or different ether residues comprising 1 to 10 carbon atoms; n is an integer ranging from 2 to 500; m is an integer ranging from 0 to 500; x is an integer ranging from 2 to 50; y is an integer ranging from 1 to 50; R same or different residue comprising a reactive group capable of forming a covalent bond with a biomolecule; F same or different fluorophores covalently bound to (S)n. The fluorescent dyes can be conjugated to a biomolecule and used for flow cytometry and/or by fluorescence microscopy.

摘要翻译： 荧光染料可以与生物分子结合并用于流式细胞术和/或通过荧光显微镜检查。

公开(公告)号：US09670318B2

公开(公告)日：2017-06-06

申请号：US14723505

申请日：2015-05-28

申请人： Miltenyi Biotec GmbH

发明人： Christian Dose

IPC分类号： C08G65/48

CPC分类号： C08G65/48 ,  A61K49/0041 ,  A61K49/0043 ,  A61K49/0045 ,  A61K49/0054 ,  A61K49/0058 ,  C08G65/33306 ,  C08G65/33396 ,  G01N33/582

摘要： The invention is directed to a fluorescent dye according to the general formula I: with C is a core moiety comprising 20 to 200 atoms; S same or different ether residues comprising 1 to 10 carbon atoms; n is an integer ranging from 2 to 500; m is an integer ranging from 0 to 500; x is an integer ranging from 2 to 50; y is an integer ranging from 1 to 50; R same or different residue comprising a reactive group capable of forming a covalent bond with a biomolecule; F same or different fluorophores covalently bound to (S)n. The fluorescent dyes can be conjugated to a biomolecule and used for flow cytometry and/or by fluorescence microscopy.

公开(公告)号：US20160298105A1

公开(公告)日：2016-10-13

申请号：US15166675

申请日：2016-05-27

申请人： Miltenyi Biotec GmbH

发明人： Jennifer BRIEDEN ,  Christian Dose

IPC分类号： C12N13/00

CPC分类号： C12N13/00 ,  G01N33/54326 ,  G01N33/569

摘要： The invention is directed to a releasable conjugate comprising a biotinylated ligand having a biotin moiety, a ligand moiety (Ligand1) and a biotin-binding molecule (bbm) bound to the biotin moiety of the biotinylated ligand. The ligand moiety of the biotinylated ligand may be separated by a spacer group consisting of polyethylene glycol. Furthermore, the invention relates to a method for cleaving the releasable conjugate by providing biotin or streptavidin and an auxiliary release agent in a sufficient concentration to displace the biotin-binding molecule (bbm) from the biotin moiety of the biotinylated ligand and a method for separation of target cells from a cell sample utilizing the conjugate.

摘要翻译： 本发明涉及包含具有生物素部分的生物素化配体，与生物素化配体的生物素部分结合的配体部分（配体1）和生物素结合分子（bbm）的可释放缀合物。 生物素化配体的配体部分可以被由聚乙二醇组成的间隔基隔开。 此外，本发明涉及通过提供足够浓度的生物素或链霉亲和素和辅助释放剂来从生物素化配体的生物素部分置换生物素结合分子（bbm）和分离方法来切割可释放缀合物的方法 的来自使用缀合物的细胞样品的靶细胞。

公开(公告)号：US20160187326A1

公开(公告)日：2016-06-30

申请号：US14944352

申请日：2015-11-18

申请人： Miltenyi Biotec GmbH

发明人： Christian Dose ,  Jennifer Brieden ,  Stefan Miltenyi ,  Thomas Rockel ,  Veronika Gann

IPC分类号： G01N33/542 ,  G01N33/569 ,  G01N33/53

CPC分类号： G01N33/542 ,  G01N33/5306 ,  G01N33/537 ,  G01N33/56972

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) Xn-P-Ym, with X is an detection moiety, P an enzymatically degradable spacer and Y an antigen recognizing moiety and n, m are integers between 1 and 100 and wherein X and Y are covalently bound to P; b) contacting the sample of biological specimens with at least one conjugate, thereby labeling the target moiety recognized by the antigen recognizing moiety Y; c) detecting the target moiety labeled with the conjugate with the detecting moiety X; and d) enzymatically degrading spacer P, thereby cleaving the detection moiety X from the conjugate. 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.

摘要翻译： 本发明涉及一种用于通过以下步骤检测生物标本样品中靶标部分的方法：a）提供至少一种具有通式（I）Xn-P-Ym的缀合物，其中X是检测部分，P酶促 可降解间隔基，Y是抗原识别部分，n，m是1〜100之间的整数，其中X和Y与P共价键合; b）使生物标本的样品与至少一种缀合物接触，从而标记由抗原识别部分Y识别的靶部分; c）用检测部分X检测用缀合物标记的靶标部分; 和d）酶促降解间隔物P，从而从缀合物切割检测部分X. 该方法可用于鉴定生物标本上的目标部分。 随后可以从样品中除去由缀合物检测的生物样品。

公开(公告)号：US11073515B2

公开(公告)日：2021-07-27

申请号：US16192805

申请日：2018-11-16

申请人： Miltenyi Biotec GmbH

发明人： Jonathan Fauerbach ,  Christian Dose ,  Thomas Rockel ,  Veronika Rudolf

IPC分类号： G01N33/543 ,  G01N33/542 ,  G01N33/53 ,  G01N33/58

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) Xn—P—Ym, b) contacting the sample of biological specimens with at least one conjugate, thereby labeling the target moiety recognized by the antigen recognizing moiety Y with the conjugate c) exciting the labelled target moieties with light having a wavelength within the absorbance spectrum of fluorescent moiety X d) detecting the labelled target moieties by detecting the fluorescence radiation and e) degrading the fluorescent moiety X of the labelled target moieties by irradiating the conjugate with light having a wavelength within the absorbance spectrum of fluorescent moiety X Use of the method in fluorescence microscopy, flow cytometer, spectrofluorometry, cell separation, pathology or histology.

公开(公告)号：US20190162721A1

公开(公告)日：2019-05-30

申请号：US16192805

申请日：2018-11-16

申请人： Miltenyi Biotec GmbH

发明人： Jonathan Fauerbach ,  Christian Dose ,  Thomas Rockel ,  Veronika Rudolf

IPC分类号： G01N33/543 ,  G01N33/542

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) Xn—P—Ym, b) contacting the sample of biological specimens with at least one conjugate, thereby labeling the target moiety recognized by the antigen recognizing moiety Y with the conjugate c) exciting the labelled target moieties with light having a wavelength within the absorbance spectrum of fluorescent moiety X d) detecting the labelled target moieties by detecting the fluorescence radiation and e) degrading the fluorescent moiety X of the labelled target moieties by irradiating the conjugate with light having a wavelength within the absorbance spectrum of fluorescent moiety X Use of the method in fluorescence microscopy, flow cytometer, spectrofluorometry, cell separation, pathology or histology

公开(公告)号：US10196631B2

公开(公告)日：2019-02-05

申请号：US15166675

申请日：2016-05-27

申请人： Miltenyi Biotec GmbH

发明人： Jennifer Brieden ,  Christian Dose

IPC分类号： C12N13/00 ,  G01N33/543 ,  G01N33/569

摘要： The invention is directed to a releasable conjugate comprising a biotinylated ligand having a biotin moiety, a ligand moiety (Ligand1) and a biotin-binding molecule (bbm) bound to the biotin moiety of the biotinylated ligand. The ligand moiety of the biotinylated ligand may be separated by a spacer group consisting of polyethylene glycol. Furthermore, the invention relates to a method for cleaving the releasable conjugate by providing biotin or streptavidin and an auxiliary release agent in a sufficient concentration to displace the biotin-binding molecule (bbm) from the biotin moiety of the biotinylated ligand and a method for separation of target cells from a cell sample utilizing the conjugate.

公开(公告)号：US20160186165A1

公开(公告)日：2016-06-30

申请号：US14944991

申请日：2015-11-18

申请人： Miltenyi Biotec GmbH

发明人： Christian Dose ,  Volker Huppert ,  Burgund Kauling ,  Philipp Steinbrück

IPC分类号： C12N13/00 ,  G01N1/40

CPC分类号： C12N13/00 ,  B03C1/01 ,  B03C1/288 ,  B03C2201/26 ,  C12N5/0087 ,  G01N1/40 ,  G01N33/54333 ,  G01N2001/4038

摘要： The invention is directed to a method for enriching target cells from a sample of cells characterized by: a) contacting the sample with a cell aggregation agent and first magnetic particles having an iron content of 0.1 pg to 5000 pg, coupled to a first antigen recognizing moiety; and second magnetic particles having an iron content of 0.05 fg to 100 fg and coupled to a second antigen recognizing moiety to obtain mixture a) b) applying a first magnetic field gradient to the mixture a) thereby removing the cells bound to the first antigen recognizing moiety coupled to the first magnetic particles, to obtain a mixture b) and obtaining an agglomerate comprising the cells of mixture a) bound to the cell aggregation agent c) applying a second magnetic field gradient to the mixture b) thereby immobilizing the cells bound to the second antigen recognizing moiety d) recovering the immobilized cells from the second magnetic field gradient as target cells.

摘要翻译： 本发明涉及用于从细胞样品中富集靶细胞的方法，其特征在于：a）使样品与细胞聚集剂和铁含量为0.1pg至5000pg的第一磁性颗粒接触，第一磁性颗粒与第一抗原识别 部分; 和铁含量为0.05fg至100fg并与第二抗原识别部分偶联以获得混合物的第二磁性颗粒a）b）向混合物施加第一磁场梯度a）从而除去与第一抗原识别结合的细胞 与第一磁性颗粒结合的部分，以获得混合物b）并获得包含与细胞聚集剂结合的混合物a）的细胞的附聚物c）向混合物施加第二磁场梯度b）从而将结合到 第二抗原识别部分d）从作为靶细胞的第二磁场梯度回收固定化细胞。

公开(公告)号：US10890580B2

公开(公告)日：2021-01-12

申请号：US15834114

申请日：2017-12-07

申请人： Miltenyi Biotec GmbH

发明人： Jennifer Pankratz ,  Christian Dose ,  Mario Assenmacher

IPC分类号： G01N33/533 ,  G01N33/536 ,  G01N1/30 ,  G01N33/569 ,  G01N33/58 ,  G01N33/532

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.