CN112585255A - 一次性生物反应器及其用途 - Google Patents
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Abstract
本发明涉及具有至少一种永久集成的一次性ATR元件的一次性生物反应器。这些生物反应器适用于培养微生物或细胞培养物，或用于无细胞的基因表达，其中通过红外光谱法检查生物反应器中的液体样品。
Description
本发明涉及具有至少一种永久集成的一次性ATR元件的一次性生物反应器。这些生物反应器适用于微生物培养、细胞培养或适用于无细胞的基因表达。通过红外光谱法对生物反应器中的样品液体进行检查。
生物反应器是在受控和监测的条件下培养不同生物的容器，例如培养用于药物制造的哺乳动物细胞。容器可由如塑料、玻璃或金属的材料制成。
如果多次使用生物反应器，则必须费力地清洁它们以使其无菌。生物反应器中的残留物会导致其他生物技术过程虚假和污染。因此，一次性生物反应器的使用更加可靠并且不易出错。
为了监测生物技术过程，可以在线(原位)测量样品。然而，对于许多测量参数，必须从生物反应器中取样，以使用单独的测量仪器进行分析。这导致只能在采样时进行测量，无法进行连续测量。对于体积非常小的生物反应器，采样会导致培养基明显减少，并增加污染的风险。
红外光谱法是一种用于对分析物进行定性和定量分析的既定分析方法，通过红外光谱法会获得表示样品化学组成特征的光谱。
红外光谱中有不同的测量原理。ATR红外光谱法是最重要一种方法。它基于衰减全反射(ATR)的原理。将分析物直接施加至ATR元件上，并将光从反面耦合至ATR元件。在ATR元件和分析物之间的界面处，一些光与样品相互作用并被样品吸收。
US9267100B2公开了一种用于生物反应器的复合传感器组件。这些传感器不是直接集成在生物反应器中，而是集成在组装的单元中。该组装的单元被胶合至生物反应器中。
从现有技术EP 3199616A1和EP 3246393A1中也已知生物反应器的一次性连接。EP3246393A1中的装置的特征在于可重新封闭的容器端口。除不同的端口外，还描述了使用由Ge、ZnSe、硫属元素化物、玻璃等材料制成的ATR元件。然而，其为连接，而不是永久集成的一次性ATR元件。
US 2010/0035337A1描述了一种带有窗口的生物反应器。该窗口的特征在于光催化涂层。描述了该窗口由ATR晶体组成。
Glindkamp等人的出版物(‘Sensors in disposable bioreactors’,Status andtrends,Adv Biochem Eng Biotechnol.2009)和Busse等人的出版物(‘Sensors fordisposable bioreactors’,Eng.Life Sei.2017,17,940-952)报道了用于生物反应器的各种传感器。认为将ATR元件集成到一次性生物反应器中是困难的，因为ATR元件的成本过高。
基于此，本发明的目的是提供一种一次性生物反应器，其中消除了现有技术中存在的上述缺点，并且允许一次性无污染的使用，同时连续确定测量参数。
具有权利要求1特征的生物反应器解决了该问题。在权利要求14中规定了根据本发明的用途。其他从属权利要求显示了其他发展。
根据本发明，提供了一次性生物反应器，其包括反应器壳体和含硅的一次性ATR元件，所述ATR元件连接至反应器壳体并且可连接至红外光谱仪。
引导红外光从光源通过一次性ATR元件至检测器。几个平行的微棱镜可以实现光的高效耦合。由于优选使用平行微棱镜，因此光学器件的定位精确度较低。
由于一次性ATR元件紧密集成，因此可以在线连续测量例如葡萄糖、乳酸盐和铵的参数。无需从生物反应器中取样，并且可以避免污染。
ATR元件优选地包括单个反射元件或由单个反射元件组成，单个反射元件优选地包括一个或多于一个微棱镜或由一个或多于一个微棱镜组成。
在另一个优选的实施方案中，ATR元件包括多个反射元件或由多个反射元件组成。
ATR元件优选是ATR红外显微镜元件。
ATR元件优选地由硅或具有涂层的硅基底组成。该涂层可以是例如由金刚石制成的薄层，或是防止细胞黏附的涂层。
ATR元件优选地具有圆形或角形。然而，任何其他形状也是可能的。
ATR元件优选气密地连接至生物反应器的侧面和/或底部。
红外光谱仪优选发出波长为2μm至20μm的光。
反应器壳体优选是气密的以防止污染。根据优选的实施方案，反应器壳体被设计为尺寸不稳定的容器。
优选反应器壳体由生物相容性聚合物制成，生物相容性聚合物特别地选自聚醚醚酮(PEEK)、聚碳酸酯(PC)、聚醚砜(PES)、聚砜(PS)、聚氯乙烯(PVC)及其组合。
优选生物反应器具有传感器，特别是用于确定生物反应器中pH或氧气含量的传感器。
如上所述，根据本发明的生物反应器用于培养微生物、细胞培养物或用于无细胞的基因表达。
基于以下附图，将更详细地解释本发明的主题，而不希望将其限于本文所示的特定形式。
图1显示了根据本发明的一次性生物反应器。
图2显示了具有集成的ATR元件的根据本发明的一次性生物反应器的详细视图。
图1显示了具有永久集成的一次性ATR元件2的一次性生物反应器1。在图中集成的一次性ATR元件作为在侧面的实例。
图2显示了具有永久集成的一次性ATR元件和所述ATR元件内的光路的一次性生物反应器的详细视图。红外辐射3通过微棱镜5耦合进出。一次性生物反应器1中的样品和一次性ATR元件2之间的界面是发生衰减全反射的地方。
附图标记的列表
·一次性生物反应器
·一次性ATR元件
·红外光
·衰减全反射
·微棱镜
Claims (14)
1.一种一次性生物反应器，其包括反应器壳体(1)和一次性的含硅的ATR元件(2)，所述ATR元件不可分离地连接至反应器壳体(1)，并且可连接至红外光谱仪。
2.根据权利要求1所述的生物反应器，其特征在于，ATR元件(2)包括单个反射元件或由单个反射元件组成，其中所述单个反射元件优选地包括一个或多于一个微棱镜(5)或由一个或多于一个微棱镜(5)组成。
3.根据前述权利要求中任一项所述的生物反应器，其特征在于，ATR元件(2)包括多个反射元件或由多个反射元件组成。
4.根据前述权利要求中任一项所述的生物反应器，其特征在于，ATR元件(2)是ATR红外显微镜元件。
5.根据前述权利要求中任一项所述的生物反应器，其特征在于，ATR元件(2)由硅组成。
6.根据权利要求1至4中任一项所述的生物反应器，其特征在于，ATR元件(2)由具有涂层的硅基底组成，所述涂层特别是薄层，例如由金刚石制成的薄层，或防止细胞黏附的涂层。
7.根据前述权利要求中任一项所述的生物反应器，其特征在于，ATR元件(2)包括圆形或角形。
8.根据前述权利要求中任一项所述的生物反应器，其特征在于，ATR元件(2)以气密方式连接至生物反应器(1)的侧面和/或底部。
9.根据前述权利要求中任一项所述的生物反应器，其特征在于，红外光谱仪发出波长为2μm至20μm的光。
10.根据前述权利要求中任一项所述的生物反应器，其特征在于，反应器壳体(1)是气密的以防止污染。
11.根据前述权利要求中任一项所述的生物反应器，其特征在于，反应器壳体(1)被设计为尺寸不稳定的容器。
12.根据前述权利要求中任一项所述的生物反应器，其特征在于，反应器壳体(1)选自聚合物，特别是聚醚醚酮(PEEK)、聚碳酸酯(PC)、聚醚砜(PES)、聚砜(PS)、聚氯乙烯(PVC)及其组合。
13.根据前述权利要求中任一项所述的生物反应器，其特征在于，生物反应器包括传感器，特别是用于确定生物反应器中pH或氧气含量的传感器，传感器优选气密地连接至反应器壳体(1)。
14.根据前述权利要求中任一项所述的生物反应器用于培养微生物或细胞培养物或用于无细胞的基因表达的用途。
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| DE102018210370.4 | 2018-06-26 | ||
| DE102018210370.4A DE102018210370A1 (de) | 2018-06-26 | 2018-06-26 | Einweg-Bioreaktor und dessen Verwendung |
| PCT/EP2019/066831 WO2020002329A1 (de) | 2018-06-26 | 2019-06-25 | Einweg-bioreaktor und dessen verwendung |
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| JPH08327533A (ja) * | 1995-06-05 | 1996-12-13 | Hitachi Ltd | 生化学分析装置 |
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Also Published As
| Publication number | Publication date |
|---|---|
| US20210115370A1 (en) | 2021-04-22 |
| WO2020002329A1 (de) | 2020-01-02 |
| CN112585255B (zh) | 2024-10-18 |
| EP3814476A1 (de) | 2021-05-05 |
| DE102018210370A1 (de) | 2020-01-02 |
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