CN108855597B - 可再充填的柱系统 - Google Patents
可再充填的柱系统 Download PDFInfo
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Abstract
本发明涉及一种用于分离颗粒的柱系统,其包括具有输入开口的储存器段;配备有过滤器矩阵和输出开口的过滤器段以及具有第一和第二端部的柱塞,第一端部装配到储存器段中,其特征在于:柱塞配备有至少一个通道,其提供从第一端部到第二端部的气体连通。用于磁性细胞分离的柱系统的使用。
Description
技术领域
本发明涉及一种用于分离颗粒的柱系统,特别是用于细胞的磁性分离。
背景技术
磁性细胞分离是一种已建立的技术,其通过利用在其中保留磁性标记的目标细胞的高梯度磁场将磁性标记的细胞与非磁性细胞分离。通常,将磁性标记的细胞和非磁性细胞的悬浮液引导通过配备有经受强磁场的铁磁性矩阵的柱体。磁性标记的细胞保留在铁磁性矩阵上,而非磁性细胞被洗提。然后可以通过从磁场去除柱体并从矩阵冲洗细胞来收获磁性标记的细胞。这种技术例如描述在文件US5693539或US5385707中。
通常在不施加压力的情况下、即在正常重力条件下执行分离过程,但是可以施加增强的压力以加速该过程或减小液体的体积。在这方面,文件US6602422公开了一种用于磁性细胞分离的微型柱系统,其配备有柱塞,其可以用于施加压力以便从柱体冲洗出液体。
理论上,用于磁性细胞分离的这样的柱体可以用于随后的若干分离过程。然而,用于磁性细胞分离的大多数柱体提供有铁磁性矩阵,如铁磁网、钢丝绒、丝绒、海绵丝或铁磁性颗粒。如果将液体从矩阵去除,例如通过排水,则空气陷入铁磁性矩阵中。由于矩阵的多孔性质,陷入的空气几乎不能被去除并且对于另外的液体而言至少部分地阻塞矩阵。这导致分离速度降低,最坏的情况是分离过程完全停止。
如在文件US6602422中公开的柱塞可用于迫使液体(和陷入的空气)通过并流出矩阵,但通过去除柱塞以添加额外的液体,空气被再次吸入到矩阵中,这次从柱体的出口端口。换言之,在US6602422中描述的柱体即使在使用了柱塞的情况下也不适合再利用。
发明内容
因此,本发明的目的是提供一种用于磁性颗粒分离的柱系统,可将压力或真空施加至该柱系统,而不存在将空气吸入或压入铁磁性矩阵中的危险。
因此,本发明的目的是一种用于分离颗粒的柱系统,其包括具有输入开口的储存器段;配备有过滤器矩阵和输出开口的过滤器段以及具有第一和第二端部的柱塞,该第一端部装配到储存器段中,其特征在于,该柱塞配备有至少一个通道,其提供从第一端部到第二端部的气体连通。
尽管可利用使用柱体的人的手指打开或闭合通道,但为了方便和安全起见,通道可以在第二端部处配备有闭合装置,如翻盖、螺帽或粘合膜,如图1或2中所示
本发明的另外的目的是一种用于分离颗粒的过程,其为柱体,其包括具有输入开口的储存器段;配备有过滤器矩阵和输出开口的过滤器段以及具有第一和第二端部的柱塞,第一端部装配到储存器段中,其中,柱塞配备有至少一个通道,其提供从第一端部到第二端部的气体连通,并具有用于位于第二端部处的通道的闭合装置,其中,将颗粒的悬浮液提供给储存器段;柱塞的第一端部在闭合装置闭合的情况下插入到柱体段的输入开口中;柱塞朝向过滤器矩阵移动,从而将悬浮液压过过滤器矩阵;打开闭合装置并从柱体去除柱塞。
可利用操作人员的手指来执行通道的打开或闭合。在备选方案中,利用提供在柱塞的第二端部处的闭合装置打开或闭合通道。
附图说明
图1和2示出了本发明的柱塞,其具有(1)通道,(2)闭合装置,(3)间隔件(垫片),(4)稳定器装置,(5)密封装置
图3示出了使用中的本发明的柱体。
具体实施方式
本发明的柱系统包括两个单独的物体:第一,包括具有输入开口(12)的储存器段(11)的柱体本身;配备有过滤器矩阵和输出开口(14)的过滤器段(13),以及第二,柱塞。
在下文中,术语“柱塞”是指类似于活塞的任何物体,其可以水密的方式插入在柱体的储存器段中。
柱塞
如图1和2中所示,在第一实施方案中,柱塞在第二端部处配备有用于通道的闭合装置(2)。适合作为闭合装置(2)的例如是翻盖、螺帽或粘合膜。
在第二实施方案中,柱塞配备有用于调节柱塞的第一端部到储存器段中的插入距离的装置(3)。距离保持器可以是圆形板(图1和2中的(3))或是从柱塞体延伸足够远以防止柱塞插入到储存器中太深的任何物体。
垫片装置或距离保持器应位于至少防止柱塞到达或接触矩阵的位置处。这确保了矩阵随时覆盖有液体并且没有空气被引入到矩阵中。优选地,柱塞在一位置处配备有垫片装置以防止柱塞插入到储存器中超过储存器的总体积的90%,优选地超过75%,且最优选地超过60%。因此,即使柱塞插入在储存器中,矩阵仍将由填充高达储存器体积的10%,25%或40%的液体覆盖。
在另一实施方案中,柱塞配备有装配到储存器段中的稳定器装置(4)。稳定器装置在图1和2中作为(4)示出,并且可提供成位于围绕柱塞的通道的翅片或拉伸壁,其从柱塞的第一端部延伸至第二端部。稳定器装置应防止柱塞的扭转和/或使机械压力能够施加至储存器中的任何液体。
柱塞、距离保持器(3)和稳定器装置(4)优选由与整体物体相同的材料制成。
柱塞的通道(1)可具有任何体积、形状或内部直径,只要它足以将空气从储存器输送到系统的外侧(即充当压力补偿)。在图2中所示的优选形式中,通道成型成双锥形(沙漏形)以向柱塞提供额外的机械稳定性。
柱塞可以在第一端部处配备有密封或垫圈(5),如O形环。没有密封的柱塞应在第一端部处具有稍大于储存器的内直径的外直径,如普通一次性注射器中的柱塞一样。具有密封的柱塞应在第一端部处具有稍小于或等于储存器的内直径的外直径。
柱体
柱体本身包括具有输入开口的储存器段;配备有过滤器矩阵和输出开口的过滤器段。只要柱塞以水密的方式装配到储存器段中,柱体的形状或体积就对本发明几乎没有影响。合适的柱体例如在文件US6602422中公开和/或可从Miltenyi Biotec私营有限责任公司以商品名“MACS柱体”得到。
柱体优选由比如聚酰胺、聚苯乙烯、聚烯烃如聚乙烯和聚丙烯、聚碳酸酯、聚甲醛、丙烯酸类如聚甲基丙烯酸甲酯、PET、聚乳酸、聚酰胺或钢等塑料制成。当矩阵用漆涂覆时,优选由将与漆结合的塑料制成,最优选的是树脂比如PCTG(用乙二醇改性的聚环己二甲基对苯二甲酸酯)。通过由亲水性材料比如亲水性塑料制造柱体可以将其制成亲水性的,或者更优选地,通过用亲水性材料例如聚乙烯吡咯烷酮涂覆柱体的内侧。
根据本发明的柱系统提供有过滤器矩阵,优选具有铁磁性矩阵。在这样的铁磁性矩阵中,通过将铁磁性矩阵所在的柱体的至少一部分插入到外部磁场中可以产生高梯度磁场。当将铁磁性矩阵带出该场时,它很容易消磁。
外部磁场可由磁轭或电磁体提供,其应提供大于约0.2特斯拉的磁场。
优选地,本发明的柱系统设计成通过重力进料来运行,但是可备选地在压力进料下运行。为了做到这一点,柱塞装备到储存器中,并且可用于利用液体冲洗柱体/矩阵。
多孔玻璃料或网格可定位在矩阵的顶端附近。多孔玻璃料/网格优选由玻璃或塑料或金属网制成,并且所具有的孔径大于或等于矩阵的孔径并小于矩阵的粒径。
铁磁性材料可包括铁磁性球或其它铁磁性颗粒。铁磁性材料可涂覆有涂层,其保持颗粒相对于彼此的相对位置。铁磁性球或颗粒优选地具有至少100µm的直径或尺寸,更优选的是大于约200µm且小于约2000µm,还更优选的是大于约200µm且小于约1000µm,并且最优选的是约280µm。
设备的制造过程
本发明的设备可通过本领域技术人员已知的任何方法来制造。优选的方法是注射成型和3D打印,例如通过挤出沉积,熔融沉积成型,立体光刻或光敏聚合物数字光处理。
本领域技术人员熟悉这样的3D打印过程和必要的器材。合适的3D打印机例如是来自Innovation MediTech私营有限责任公司的具有FotoMed® LED.A作为印刷光刻胶聚合物的M120 Scan-LED打印机。通常,具有25µm厚度的聚合物的层通过UV辐射固化,随后通过摇晃去除未固化的液体FotoMed® LED.A。在将最后一层印刷/固化之后,通过在填充有异丙醇的超声波浴中冲洗物体来去除未固化的FotoMed® LED.A。最后,可以执行在N2氛围中,例如在后固化单元PCU90(Innovation MediTech私营有限责任公司)中的后固化,。
设备的使用
通常,本发明的过程涉及分离颗粒。尤其,该过程涉及细胞的分离(即颗粒是细胞),并且通过将细胞磁性标记来进行分离,并通过将柱体放置在磁场中来将磁性标记的细胞保留在矩阵中。
因此,在优选的实施方案中,过滤器矩阵是铁磁性矩阵,并且该过程至少部分地通过将柱体的过滤器段放置在磁场中来执行。
将细胞磁性标记的各种技术在本领域已知超过20年。这些过程的共同之处在于经由与磁性颗粒偶联的抗体对细胞进行选择性的磁性标记,将细胞作为悬浮液引入到放置在强磁场中的柱体中,并将标记的细胞保留在矩阵上。为了在分离之后从柱体获得磁性标记的细胞,通常用缓冲液冲洗矩阵,并且因此也将空气冲洗到矩阵中。由于矩阵中的空气,柱体不再起作用,并且不可能将其再用于进一步的分离。
利用本发明的柱体,可在不用空气冲洗柱体的情况下洗提目标材料。在图3中显示了本发明的过程,在其中将缓冲液添加至柱体,并且通过将具有闭合的盖的柱塞推动到柱体中,将目标材料从柱体洗提,但矩阵仍覆盖有液体如缓冲液。
垫片防止柱塞插入到储存器(a)中太深。然后,通过打开闭合装置(b)可以将柱塞从储存器去除而不会将空气通过出口(c/d)吸入到柱体的矩阵中。矩阵不包含任何陷入的空气,矩阵仍然是湿润的并且可以用于进一步的分离过程。
Claims (9)
1.用于分离磁性颗粒的柱系统,其包括具有输入开口的储存器段;配备有过滤器矩阵和输出开口的过滤器段以及具有第一和第二端部的柱塞,所述第一端部装配到所述储存器段中,并且所述过滤器矩阵是铁磁性矩阵,其特征在于,所述柱塞配备有至少一个通道,其提供从所述第一端部到所述第二端部的气体连通。
2.根据权利要求1所述的柱系统,其特征在于,所述通道在所述第二端部处配备有闭合装置。
3.根据权利要求1或2所述的柱系统,其特征在于,所述柱塞配备有用于调节所述柱塞的第一端部到所述储存器段中的插入距离的装置。
4.根据权利要求1或2所述的柱系统,其特征在于,所述柱塞配备有装配到所述储存器段中的稳定器装置。
5.根据权利要求1或2所述的柱系统,其特征在于,所述柱塞在第一端部处配备有密封装置。
6.根据权利要求1或2所述的柱系统,其特征在于,所述柱塞配备有作为闭合装置的翻盖、螺帽或粘合膜。
7.用于在柱体中分离磁性颗粒的过程,所述柱体包括具有输入开口的储存器段;配备有过滤器矩阵和输出开口的过滤器段以及具有第一和第二端部的柱塞,所述第一端部装配到所述储存器段中,并且所述过滤器矩阵是铁磁性矩阵,其中,所述柱塞配备有至少一个通道,其提供从所述第一端部到所述第二端部的气体连通,其特征在于,将颗粒的悬浮液提供给所述储存器段;所述柱塞的第一端部在所述通道闭合的情况下插入到所述柱体段的输入开口中;所述柱塞朝向所述过滤器矩阵移动,从而将所述悬浮液压过所述过滤器矩阵;打开所述通道并将所述柱塞从所述柱体去除。
8.根据权利要求7所述的过程,其特征在于,所述通道利用在所述柱塞的第二端部处提供的闭合装置来打开或闭合。
9.根据权利要求7或8所述的过程,其特征在于,所述过程至少部分地通过将所述柱体的过滤器段放置在磁场中来执行。
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