CN101842690B - 受抑全内反射生物传感器盒 - Google Patents

受抑全内反射生物传感器盒 Download PDF

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CN101842690B
CN101842690B CN2008801136389A CN200880113638A CN101842690B CN 101842690 B CN101842690 B CN 101842690B CN 2008801136389 A CN2008801136389 A CN 2008801136389A CN 200880113638 A CN200880113638 A CN 200880113638A CN 101842690 B CN101842690 B CN 101842690B
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J·H·尼乌文赫伊斯
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Abstract

本发明提供了一种生物传感器盒，其包括具有适于容纳液体样品的孔室的底部以及用于封闭所述孔室的盖部。所述孔室具有传感器表面。所述底部适于允许光沿第一光学路径进入，在所述传感器表面被反射，以及沿第二光学路径离开。本发明还涉及一种制造此类盒的方法。

Description

受抑全内反射生物传感器盒
技术领域
本发明涉及一种用于受抑全内反射(FTIR)生物传感器的盒以及一种制造此类盒的方法。 
背景技术
目前，对生物传感器的需求日益增长。通常，生物传感器允许检测分析物中的给定特异性分子，其中，所述分子的量通常很小。例如，人们可以测量唾液或血液内的药物或心血管疾病标记物的量。因此，如果分析物中存在待检分子，那么可以使用仅与特异性结合部位或结合点结合的靶粒子，例如超顺磁标记珠。一种用于检测这些结合到结合点的标记粒子的已知技术是FTIR。其中，将光以全内反射的角度与样品耦合。如果靠近样品表面没有粒子，那么光将被完全反射。然而，如果标记粒子被结合到所述表面，那么全内反射的条件被妨碍，一部分光散射进入样品，因而减小了被所述表面反射的光的量。通过使用光学检测器测量反射光的强度，可以估计结合到所述表面的粒子的量。这使得可以估计存在于分析物或样品中的感兴趣的特异性分子的量。 
由于该技术被认为将成为生物感测中的标准工具，因此对于可以用于FTIR的盒的需求日益增长。由于基于免疫反应的生物传感器因为盒内的生化材料在试验期间会改变而需要是一次性的，因此这种情况下尤其需要便宜的用于FTIR的一次性盒或其他检测技术。 
US 2005/0179901A1公开了一种检测器装置，其包括可旋转的微流控盘和分光光度检测器单元。该装置基于SPR并且能够测量检测微腔内的分析物，每个检测微腔是部分的微通道结构。 
EP0949002A2公开了一种用于制造具有支撑层的分析支撑装置的方法，在所述支撑层上层压有距离保持器层。 
US4849340公开了一种用于方便执行液体测定的元件和方法。所述元 件使用毛细管作用来将预定体积的液体样品吸入装满试剂的反应室中，在该反应室中监测液体样品与试剂之间的反应。 
US6607701B1公开了一种用于连续生产一次性毛细管塑料微量吸收池的方法，其包括如下步骤：提供第一片状物和第二片状物，提供至少一个低压使得在所述片状物的至少一个中的预定地方有预定深度，以至少一个低压引入至少一种添加剂或试剂，连接所述第一片状物和第二片状物以获得包括至少一个腔的体构件。 
发明内容
因此，本发明的目的是提供一种用于生物传感器的改进的盒。该目标由独立权利要求的特征实现。 
本发明提供了一种用于生物传感器的盒，其包括具有适于容纳样品的孔室的底部，以及用于封闭所述孔室的盖部，所述孔室具有传感器表面，其中，所述底部适于允许光沿第一光学路径进入，在所述传感器表面处被反射，并沿第二光学路径离开，其中，所述盖部包括带或箔。任选地，可以在底部和盖部之间提供粘结剂层。这可能是有利的，尽管粘结剂层在所述孔室处或上方被中断。因此，阻止了孔室内的样品和粘结剂层之间的任何相互作用。 
本发明也提供一种制造盒的方法，包括如下步骤： 
a)提供具有适于容纳样品的孔室的由第一材料制成的底部； 
b)提供由第二材料制成的卷形物或片状物； 
c)将试剂和/或标记粒子施加到卷状物或片状物的特异性部位； 
d)将所述卷状物或片状物裁剪为盖部，其中，每个盖部包含所述试剂和/或标记粒子；以及 
e)将所述盖部附接到所述底部，这样试剂和/或标记粒子被封闭在所述孔室中。 
所述盒的底部可以由塑料制成，优选地塑模成形。任选地，可以在底部和盖部之间提供粘结剂层。这将是有利的，尽管所述粘结剂层在所述孔室上或上方被中断。因此，阻止了在孔室内的样品和粘结剂层之间的任何相互作用。 
在优选的实施例中，所述孔室包括道。所述孔室可以具有比所述道更大的宽度和深度，但也可以预见到整个孔室由道组成。所述盒还包括用于向所述孔室中提供样品的至少一个管道或开口。所述管道或开口可以是，例如，底部中的槽或盖部中的孔。也可以预见到的是，所述道在一端或两端开口。 
根据优选实施例，所述盒的底部包括适于容纳用于提供磁场的装置(例如线圈)的凹进处。此外，所述底部可以包括分别在第一光学路径内的光学输入表面和在第二光学路径内的光学输出表面。优选地，这些表面垂直于所述第一光学路径和第二光学路径。 
此外，所述孔室可以包含试剂或若干种试剂的组合以及标记粒子。在优选实施例中，这些试剂位于所述传感器表面的特异性结合点。根据另一实施例，在盖部上提供试剂和/或标记粒子。例如，它们可以印刷到形成盖部的薄片上。 
所述标记粒子可以涂布有捕获分子并且还可以包括磁粒子。例如，所述标记粒子若是超顺磁性的将是有利的。 
本发明还涉及一种制造上述盒的方法。根据所述方法，提供了具有适于容纳样品的孔室的由第一材料制成的底部。此外，提供了由第二材料制成的卷形物或片状物。随后，将试剂和/或标记粒子施加到在所述卷形物或片状物上的特异性部位，以及将所述卷形物或片状物切裁剪为盖部，其中，每个盖部包含所述试剂和/或标记粒子。最终，将所述盖部附接到所述底部，使得所述试剂和/或标记粒子被封闭在所述孔室中。任选地，所述方法可以包括将粘结剂层应用到所述底部和/或由第二材料制成的卷形物或片状物的步骤。 
本发明的这些或其他方面将从下文所描述的实施例中变得显然，以及将参考这些实施例进行阐述。 
附图说明
图1a示例性地示出了根据本发明的盒的横截面； 
图1b示意性地示出了根据本发明的所述盒的底部的顶视图； 
图1c示意性地示出了根据本发明的另一盒的底部的顶视图； 
图2示意性地示出了FTIR的功能原理； 
图3a示意性地示出了用作盖部的材料的卷形物； 
图3b示意性地示出了用作盖部的材料的片状物； 
图4示意性地示出了所述盒的替代结构的侧视图。 
具体实施方式
图1a示意性地示出了作为本发明的实例的盒11的横截面的侧视图。盒11包括具有孔室2的底部1以及盖部3。孔室2适于填充样品，并且由盖部3封闭或覆盖。传感器表面4限定出孔室2的下底面。光沿第一光学路径5在光学输入表面5a进入底部1，在所述传感器表面4处被反射，并且沿第二光学路径6在光学输出表面6a离开底部1。底部1还形成有凹进处7，所述凹进处7适于容纳用于提供磁场的装置，然后所述磁场对磁标记粒子8施力。这样做是为了实现试剂和标记粒子8之间的结合，即所谓的致动过程。孔室2中包含试剂(未示出)和标记粒子8。 
如图1b中可以看到的，该附图示意性地示出盒11的底部1的顶视图，孔室2可能还包括道9。整个孔室2也可能由道9构成，如图1c中所示。在末端，道9包括用于向孔室2提供样品的管道或开口10。例如，所述管道或开口10可以是图1b和1c中所描绘的底部的槽，或者盖部3上的孔。 
因此对孔室2的填充可能通过毛细管力实现：若将道9的直径选择为足够小，则在小滴样品接触管道或开口10时，它将被吸入孔室2。在这种情况下，显然需要提供第二管道或开口10以允许空气离开孔室2。然而，也可以通过使用例如注射器等施加压力来填充孔室2。 
图2示意性地示出了作为盒11所使用的检测方法的实例的FTIR的功能原理。一旦孔室2被填充或被提供了如上所描述的待检液体样品，以干物质形式提供的标记粒子8扩散到溶液中。使用磁体13，超顺磁标记粒子8可以向传感器表面4加速，在传感器表面4处，若样品中存在待检特异性分子，则这些标记粒子可以结合到所述表面上。在足够进行结合的时间之后，为了从所述表面去除没有结合到传感器表面14上的标记粒子8，可以使用磁体14。在该“冲洗”步骤之后，使用激光或LED 16照射传感器表面4。光在传感器表面4处被反射并被检测器12检测，所述检测器12可以 是光电二极管或CCD照相机。通常，在测定过程期间、上述的结合过程期间连续读出光学元件或检测器12，并且监测所述结合过程的进程。然而，或者可以获得测定之前的图像和测定之后的一幅图像，然后比较两者的差别。选择入射光的光学路径5以便满足全内反射的条件。在这种情况下，产生光学倏逝场，所述光学倏逝场通常仅穿透50-100nm进入孔室2。因此，只要标记粒子8靠近传感器表面4，所述倏逝场被扰乱并导致反射强度的降低。 
本发明还涉及一种制造上述盒11的方法。根据所述方法，提供了具有适于容纳样品的孔室2的由第一材料制成的底部1。所述第一材料优选是塑料材料，例如聚苯乙烯、聚碳酸酯、环烯烃聚合物(COP或ZeonexTM)或PET(聚对苯二甲酸乙二酯)。底部1可以使用所述第一材料塑模成形，例如，注塑成型。然而，也可以使用形成底部1的其他技术。 
此外，提供由第二材料制成的卷形物15或片状物18，如在描绘出卷形物15的图3a中和在描绘出片状物18的3b中所示。所述卷形物15或片状物18可以包括由例如聚酯制成的箔或带。如果卷形物15或片状物18包括箔，那么可以施加另一粘结剂层以将箔粘附到盒11上。在某些实施例中，所述粘结剂可以仅仅部分覆盖卷形物15或片状物18。 
随后将试剂和/或标记粒子8施加到卷形物15或片状物18的特异性部位，并且将卷形物15或片状物18裁剪为盖部3，其中，每个盖部包含试剂和/或标记粒子8。最终，将盖部3附接到盒11的底部1，使得所述试剂和/或标记粒子8被封闭在孔室2中。如果在卷形物15或片状物16上没有粘结剂，也可以使用其他结合技术将盖部3附接到底部1，例如，可以使用诸如超声波焊接和激光焊接的热结合技术。 
或者，可以将试剂提供给孔室2的传感器表面4，特别地，优选为传感器表面4上的若干特异性结合点提供试剂。不同结合点的试剂也可以彼此不同，以便向待分析样品中的不同分子提供特异性结合点。这些分子例如可以是抗体或共轭分析物分子(例如，在抑制测定的情况下)。 
图4示出了类似于图1a的盒11的实例的另一示意性横截面侧视图。底部1类似于图1a的底部1，例如，由塑料塑模成形。提供用于沿光学路径5输入光的光学输入表面5a以及用于沿光学路径5输出光的光学输出表 面6。在底部1中形成的孔室2被盖部3覆盖，由此内嵌形成封闭的空间。图4的实例与图1a的实例的差别在于，传感器表面4不是基本平的表面，而是具有锯齿状的结构。在传感器表面4处提供至少一个楔形结构，导致光有不同路径撞击传感器表面4。通常，光的路径然后是在下倾面的反射，因而大多数光穿过所述面，所述光被反射到相对的上倾面，从而传感器表面4的下倾面和上倾面形成像等腰三角形等边的凹进处。从第二下倾面，所述光再次被反射，沿图4所示的第二光学路径6向着光学输出表面6a的方向离开孔室2。具有试剂和/或标记粒子8(此处未示出)的待检样品，再次位于传感器表面4，基本上位于楔形棱面之间。检测原理是光基本上被结合到所述样品的分子上的标记粒子8阻挡。通过计算沿第一光学路径5的入射光与沿第二光学路径6的出射光的差别，能够推断样品中分子的量。 
本发明重要的一方面是，如所描述的，盒11是由包括用于包含样品的孔室2的底部1和覆盖孔室2的单一盖部3构造的，所述盖部3是带或箔。该结构使得盒11便宜并且可以用作生物传感器中的一次性部分。就此而言，带被定义为一侧有粘性，并且由不同的常用材料形成为的扁平柔性体。就此而言，箔被定义为不不具有粘性，并且由不同的常用材料形成为的扁平柔性体。在后一种情况下，在底部1和箔之间提供粘结剂层。带和箔以在底部1的边缘被裁剪的条状应用于底部1。 
虽然在附图和说明书中已经详细举例说明和描述了本发明，但是此举例说明和描述应当被理解为是举例说明性和示例性的，而不应是限制性的；本发明并不限于所公开的实施例。本领域技术人员在根据对附图、说明书和权利要求书的研究来实践所要求的发明时能够想到并实现所公开的实施例的其他变型。在权利要求书中，词语“包括”并不排除其他元件或步骤，并且不定冠词“一”或“一个”并不排除多个。单一处理器可以实现权利要求书中记载的若干项的功能。起码的事实是，在相互不同的权利要求中记载的特定手段并不代表使用这些手段的组合不能带来益处。权利要求中的任何附图标记不能解释为是对范围的限制。 

Claims (15)

1.一种用于生物传感器的盒（11），包括具有适于容纳样品的孔室（2）的底部（1），以及用于封闭所述孔室（2）的盖部（3），所述孔室（2）具有传感器表面（4），其中，在所述传感器表面（4）处提供至少一个楔形结构，并且所述样品位于所述传感器表面（4）的楔形棱面之间，其中，所述底部（1）适于允许光沿第一光学路径（5）进入，在所述传感器表面（4）处被反射，以及沿第二光学路径（6）离开，其中，所述盖部（3）包括带或箔。
2.如权利要求1所述的盒（11），其中，所述底部（1）由塑料制成和/或所述底部（1）是塑模成形的。
3.如权利要求1所述的盒（11），其中，所述孔室（2）包括道（9），并且所述孔室（2）包含试剂或若干种试剂的组合和/或标记粒子（8），其中，所述试剂或若干种试剂的所述组合位于所述传感器表面（4）的特异性结合点。
4.如权利要求1所述的盒（11），还包括在底部（1）和盖部（3）之间的粘结剂层，其在所述孔室（2）处被中断。
5.如权利要求3所述的盒（11），其中，所述标记粒子（8）涂布有捕获分子。
6.如权利要求5所述的盒（11），其中，所述标记粒子（8）包括磁性粒子。
7.如权利要求1所述的盒（11），还包括用于向所述孔室（2）提供所述样品的至少一个管道或开口（10）。
8.如权利要求7所述的盒（11），其中，所述至少一个管道或开口（10）是在所述底部（1）中的槽。
9.如权利要求7所述的盒（11），其中，所述至少一个管道或开口（10）是在所述盖部（3）中的孔。
10.如权利要求1所述的盒（11），其中，所述底部（1）包括用于容纳用于提供磁场的装置（13）的凹进处（7）。
11.如权利要求1所述的盒（11），其中，所述底部（1）包括分别在第一光学路径（5）内的光学输入表面（5a）和第二光学路径（6）内的光学输出表面（6a）。
12.如权利要求11所述的盒（11），其中，所述表面（5a、6a）垂直于所述光学路径（5、6）。
13.如权利要求1所述的盒（11），其中，所述盖部（3）包括具有产品信息和/或标识的印刷标记。
14.一种制备如权利要求1所述的盒（11）的方法，包括如下步骤：
a）提供具有孔室（2）的由第一材料制成的底部（1），所述孔室（2）具有提供有至少一个楔形结构的传感器表面（4），所述孔室（2）适于在所述传感器表面（4）的所述楔形棱面之间容纳样品；
b）提供由第二材料制成的卷形物（15）或片状物（18）；
c）将试剂和/或标记粒子（8）施加到所述卷形物（15）或片状物（18）上的特异性部位；
d）将所述卷形物（15）或片状物（18）裁剪为盖部（3），其中，每个盖部（3）包含所述试剂和/或标记粒子（8）；以及
e）将所述盖部（3）附接到所述底部（1），使得所述试剂和/或标记粒子（8）被封闭在所述孔室（2）中。
15.如权利要求14所述的方法，还包括将粘结剂层施加到所述底部（1）和/或由第二材料制成的所述卷形物（15）或片状物（8）的步骤。
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US8449833B2 (en) 2013-05-28
BRPI0818571A2 (pt) 2015-04-22
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RU2497100C2 (ru) 2013-10-27
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