CN1846131B - 制备窄的均匀试剂条的方法和试剂 - Google Patents

制备窄的均匀试剂条的方法和试剂 Download PDF

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CN1846131B
CN1846131B CN2004800239248A CN200480023924A CN1846131B CN 1846131 B CN1846131 B CN 1846131B CN 2004800239248 A CN2004800239248 A CN 2004800239248A CN 200480023924 A CN200480023924 A CN 200480023924A CN 1846131 B CN1846131 B CN 1846131B
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C·D·维尔西
D·莫索伊乌
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T156/00—Adhesive bonding and miscellaneous chemical manufacture
    • Y10T156/10—Methods of surface bonding and/or assembly therefor
    • Y10T156/1052—Methods of surface bonding and/or assembly therefor with cutting, punching, tearing or severing
    • Y10T156/1059—Splitting sheet lamina in plane intermediate of faces
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T156/00—Adhesive bonding and miscellaneous chemical manufacture
    • Y10T156/10—Methods of surface bonding and/or assembly therefor
    • Y10T156/1052—Methods of surface bonding and/or assembly therefor with cutting, punching, tearing or severing
    • Y10T156/1084—Methods of surface bonding and/or assembly therefor with cutting, punching, tearing or severing of continuous or running length bonded web

Abstract

本发明涉及可以在制备测试条的工艺中在模缝涂覆平整的支撑材料中使用的试剂涂层体。有利的是，本发明的试剂体显示出某些优异的流变性性质，比如粘度、表面张力和触变性。该试剂体优选用来在平整的薄片材料上涂覆薄、窄而均匀的试剂材料条。

Description

制备窄的均匀试剂条的方法和试剂
相关申请 
本申请要求2003年6月20日提交的系列号为No.60/480397的美国临时专利申请的优选权。本申请和与本申请同一日期提交的共同受让申请“Reagent Stripe for Test Strip”(下面记为“试剂条申请”)相关，所述共同受让申请在此全文引入作为参考。 
发明领域
本发明一般涉及生物传感器中使用的试剂，或者测试条，更具体涉及在测试条的平整表面上制备窄的均匀试剂条. 
背景和综述 
在众多向测试条上施加试剂的方法中，电化学生物传感器过去主要通过采用印刷技术比如丝网印刷工艺或者分配技术来施加液体试剂并随后干燥进行制备(参见例如美国专利No.5437999和WO97/02487)。这些分配方法已经成功用于所谓的“毛细填充”测试条，比如Roche Diagnostics  
Figure DEST_PATH_G13621204150131000D000011
Advantage测试条的制备。尽管这些技术可以制备可靠的电化学生物传感器，但是对于高产量生产线并不太适用。另外，这些分配技术受到试剂干燥不均匀这个缺点的困扰，导致电极的覆盖区域上试剂厚度不均一。而且，上述技术不适于可靠地、可重复性地制备超薄地试剂层(10微米或更薄)。所以，需要有改进的试剂施加工艺。 
在制备涂覆在例如透明聚合物基体上的试剂薄膜时(例如，美国专利No.5437999和6036919)，人们已经建议并成功采用了刮涂将试剂组合物涂覆到平整的基体上。通常，采用这种工艺可以制备宽度为数厘米到数米的薄膜。为了制备测试条，将这样涂覆的试剂层切成小条，然后施加到测试条基体上.刮涂试剂体的缺点在于——尽管薄膜的中心部分厚度均匀——但涂覆区域的边缘发现有不均匀现象，相信这是源于干燥效应和边缘效应。尽管在基体上涂覆宽试剂带的情况下，由于涂层的边缘不均匀部分可以通过边缘修整而去除，使得这些不均匀现象可以接受，但是随着待涂覆的试剂条变小/窄，这些不均匀现象越来越不能接受。 
WO02/057781公开了由薄膜材料制备试剂条的方法。其中，它公开了可以通过敷设窄条试剂材料而将该试剂材料施加到条支撑材料上，其中所述窄条试剂材料可以由或不由支撑载体支撑。 
美国专利申请公开2003/0097981、美国专利公开No.2003/0099773、美国专利No.6676995和6689411和EP1316367公开了用于将试剂溶液条敷设到基体上的溶液剥离系统(solutionstripping system)。该系统使得试剂溶液可以模缝涂覆到薄片材料上，例如用于电化学葡萄糖传感器，所述溶液粘度低，为约0.5-25厘泊(cP＝mPa-s)。 
美国专利No.3032008、3886898和4106437教导了可用来将液体材料涂覆到固体支撑上的涂覆装置。 
美国专利No.6036919公开了用于光学血液葡萄糖测试条的试剂薄膜。该试剂组合物包括黄原胶和其它物质。 
美国专利申请公开No.2003/0146113公开了用于电化学凝结传感器的试剂薄膜。该试剂组合物包括充当薄膜形成体的羧酸酯化微晶纤维素( 
Figure S04823924820060303D000021
R591)，和其它物质。 
上述参考文献都不能满足在固体支撑材料上可靠地形成窄的(例如，小于1cm)、薄的(例如，小于10μm)和均匀的试剂条以用于制备测试条、特别是电化学测试条的方法的要求。 
所以，本发明的目标是提供一种方法和相应的试剂组合物，利用该方法和试剂组合物，可以在例如薄片材料的平整表面上，特别是电化学生物传感器测试条的电极区域上，沉积超薄、窄而且均匀的试剂线或条。 
本目标通过本发明得以实现，其中本发明涉及用于制备窄而均匀的试剂条的模缝涂覆工艺的试剂。 
第一方面，本发明涉及显示出剪稀性、轻微触变性的或者触变性行为的试剂组合物。 
第二方面，本发明涉及采用模缝涂覆工艺将该剪稀性的、轻微触变性的或触变性试剂组合物涂覆到薄片材料上的方法。 
在又一方面，本发明涉及包含该剪稀性、轻微触变性或触变性试剂的分析测试元件。 
在另一方面，本发明涉及具有剪稀性而且至少轻微触变性的试剂 组合物。它还涉及分析测试元件以及制备包括该剪稀性而且轻微触变性的试剂组合物的分析测试元件的方法。 
附图简述 
图1以6个步骤(A-F)示意性给出了如何采用本发明的模缝涂覆工艺制备具有单一试剂区域的电化学测试元件。 
图2以6个步骤(A-F)示意性给出了如何采用本发明的模缝涂覆工艺制备具有两个试剂区域的电化学测试元件。 
图3给出了横跨实施例1的试剂条测量的表面光洁度测量结果。 
图4示出了横跨根据本发明的试剂条测量的表面光洁度测量数据。 
图5示出了横跨不使用本发明的流变改性剂的试剂条的表面光洁度测量数据。 
图6是本发明的涂覆在薄片材料上的试剂条的显微照片。 
图7给出了涂覆在薄膜材料上的不使用本发明的流变改性剂的试剂条的两张显微照片(图7A和7B)。 
优选实施方案描述 
为了帮助理解本发明的原理，现在请参见本文示例的具体实施方案，而且将采用具体的语言来描述这些具体实施方案。然而应该理解，由此并不意在限制本发明的范围。对所述工艺或设备的任何改变和进一步修改，以及对本文所述本发明的原理的任何进一步应用，都认为对本发明所涉及领域的技术人员而言是正常情况下会发生的。本发明的优选实施方案是从属权利要求的主题。 
本发明的试剂组合物是剪稀性的、轻微触变性的或者触变性的。触变性试剂组合物是显示出流变行为的试剂组合物，具体取决于在该试剂组合物上是否施加了外部剪切力。剪稀性试剂组合物是当在其上施加了剪力时变稀，即粘度变小的试剂组合物。通常，在向本发明的试剂组合物施加剪力之前，该组合物具有一定的粘度。在施加剪力时，该组合物粘度下降。如果终止剪力后粘度再次增加——具有一定的时间相关性，则认为该试剂组合物是“剪稀性的”。如果终止剪力后粘度增加仅仅具有一定的延迟性，则认为该试剂组合物是“触变性的”。 
触变性是假塑性的特殊例子。触变性流体经历“剪稀行为”。但是，随着剪力下降，粘度发生重建，以较慢的速率增加，从而产生滞 后回线。轻微触变性的流体的滞后回线不太明显。另外，触变行为显著受到所研究材料的剪切历史的影响。在对比性测量中，应该小心以确保待比较的试样具有相同或者至少非常相似的历史。 
本发明的试剂组合物可用于模缝涂覆工艺。在模缝涂覆中，通过迫使试剂液体或浆料通过模缝涂布头的狭缝，将该流体试剂组合物施加到固态基体上，优选薄片材料形式的基体上。通常，薄片材料以一定速度通过狭缝一定距离。但是，也可以是模缝涂布头横跨该薄片材料移动，或者是模缝涂布头和薄片都移动。 
为了实现本发明的目标，有利的是用作涂料体的试剂组合物的流变性质位于一定的优选范围内：粘度优选是约70-约130mPa-s，最优选为95-115mPa-s。有利的表面张力范围是30-50mN/m，优选是约40±2mN/m。该涂料体显示出剪稀性、轻微触变性或触变性行为也很重要。 
本发明的一个方面是在该试剂涂覆体中掺入黄原胶。可以使用的一种黄原胶品牌是 
Figure S04823924820060303D000041
。该组分对试剂体的触变性有影响。含有黄原胶的试剂涂覆体，比如 
Figure S04823924820060303D000042
，允许制备超薄的试剂层。优选地，试剂层干燥薄膜的厚度小于10μm，干燥试剂层的特别优选厚度为1.5-5μm。 
已经证明，在本发明的试剂组合物中结合二氧化硅对于该试剂的粘度和触变性有有利的影响。通过添加二氧化硅，这两种性质都得到了改善。优选采用未处理的亲水性二氧化硅。优选形式的二氧化硅的颗粒尺寸为约1-7μm。已经证明，二氧化硅出乎意料地改善了涂覆体的其它组分，尤其是羧甲基纤维素和 ，的触变行为。另外，干薄膜中的二氧化硅颗粒防止涂覆的条和薄片背面之间发生背面传递，从而使得涂覆的薄片材料可以以材料卷的形式存储。另外，干薄膜中的二氧化硅颗粒增加了试剂涂层的比表面积，使得例如试剂可以快速溶解在试样液体中。在包括含有本发明试剂组合物的试剂条的毛细填充生物传感器中，二氧化硅还改善了毛细填充时间并促进了组分在试剂条中的迁移。 
另一种改善试剂粘度和触变性的添加剂是羧甲基纤维素(CMC)。所以，本发明试剂组合物的特别优选实施方案包括黄原胶例如 
Figure S04823924820060303D000044
二氧化硅和CMC。 
本发明的试剂组合物使得可以形成薄的试剂层，例如，制备电化学生物传感器。薄试剂层有多个优点： 
和试剂组分相比，试样组分过量，从而不会限制决定性反应(determination reactions)。 
薄试剂层可以制备成均匀的厚度。 
薄试剂层仅仅含有少量试剂，这进而导致反应时间快。 
反应仅仅具有短的扩散时间。 
薄试剂层可以快速溶解，所以导致在试剂条发生试样再水化后可以很快使用试剂而且基质很快达到平衡，这进而使得可以快速测量。 
本发明的试剂层不仅仅可以制备成非常薄，而且在反应区的薄片纵向和横向都显示出高度的均匀性。试剂层在测试区域平整而且厚度均匀。涂覆的条的厚度变化优选仅仅发生在条的外围0.2cm(或更少)边缘。在优选实施方案中，有利的做法是这些区域可以或者在传感器组装期间由分隔层覆盖，或者可以在最后组装工艺中从完成的传感器中修整掉。 
除了影响本发明试剂组合物的流变学性质的上述组分以外，试剂还可以包括下列物质类中的一种或多物质(成分)。可以加到试剂中的物质、添加剂和成分包括但不限于下列： 
缓冲液，例如，硫酸盐缓冲液； 
酶，比如，葡萄糖脱氢酶、葡萄糖染料氧化还原酶、葡萄糖氧化酶和其它氧化酶或脱氢酶比如用于测定乳酸盐或者胆固醇、酯酶等； 
介体，比如亚硝基苯胺类、铁氰化物、六胺钌、锇络合物； 
稳定剂，比如海藻糖、琥珀酸钠； 
增稠剂，比如 
Figure S04823924820060303D000051
、CMC 
蛋白质，比如酶、牛血清蛋白 
指示剂； 
染料； 
表面活性剂，比如 
Figure S04823924820060303D000052
Figure S04823924820060303D000054
DONS； 
成膜剂，比如 
Figure S04823924820060303D000056
聚乙烯吡咯烷酮、聚乙烯醇、 
Figure S04823924820060303D000057
酶的辅因子，比如NAD、NADH、PQQ；和 
二氧化硅，例如，DS300、DS320、DS300的球磨二氧化硅、DS320的球磨二氧化硅。 
下表1列出了可用于测量特殊分析物的酶和介体的非限制性实施例。 
表1是可用来测量特定分析物水平的一些分析物、酶和介体的部分列表。 
  分析物   酶  介体(氧化形式)   辅助介体
  葡萄糖   葡萄糖脱氢酶和  心肌黄酶  铁氰化物  
  葡萄糖   葡萄糖脱氢酶  (喹啉蛋白质  (quinoprotein))  铁氰化物  
  胆固醇   胆固醇酯酶和胆  固醇氧化酶  铁氰化物   2，6-二甲基-1，4-苯醌  2，5-二氯-1，4-苯醌或者吩嗪乙基  硫酸盐(ethosulfate)
  HDL  胆固醇   胆固醇酯酶和胆  固醇氧化酶  铁氰化物   2，6-二甲基-1，4-苯醌  2，5-二氯-1，4-苯醌或者吩嗪乙基  硫酸盐(ethosulfate)
  甘油三  酸酯   脂蛋白脂肪酶、  甘油激酶和甘油-  3-磷酸酯氧化酶  铁氰化物或吩嗪乙基硫 酸盐(ethosulfate)   吩嗪甲基硫酸盐(methosulfate)
  乳酸酯   乳酸酯氧化酶  铁氰化物   2，6-二甲基-1，4-苯醌
  乳酸酯   乳酸酯脱氢酶和  心肌黄酶  铁氰化物吩嗪乙基硫酸 盐(ethosulfate)、或者吩 嗪甲基硫酸盐 (methosulfate)  
  乳酸酯  脱氢酶   心肌黄酶  铁氰化物   吩嗪乙基硫酸盐(ethosulfate)、或  者吩嗪甲基硫酸盐(methosulfate)
  丙酮酸  酯   丙酮酸酯氧化酶  铁氰化物  
  乙醇   乙醇氧化酶  亚苯基二胺  
  胆红素   胆红素氧化酶  1-甲氧基-吩嗪甲基硫酸 盐(methosulfate)  
  尿酸   尿素酶  铁氰化物  
在表1所示的一些实施例中，至少一种辅助酶用作反应催化剂。另外，表1所示的一些实施例可以采用辅助介体，该辅助介体便于电子传递到介体的氧化形式。该辅助介体可以以比介体的氧化形式少的量提供给试剂。尽管描述了上面的试验，但可以设想，采用根据本公开的电化学传感器，试样的电流、电荷、电阻、电导、电势或其它电化学指示的性质可以和试样中分析物的浓度精确相关。 
实施例1、2、3和4给出了分别针对电化学血葡萄糖和凝结传感器的试剂组合物的例子。 
在优选实施方案中，上述试剂组合物施加到基体上，所述基体已经通过模缝涂覆工艺含有电化学传感器的电极迹线(electrode trace)或线路。实施例5给出了这种工艺的例子。 
这些电极线路的优选制备技术采用激光蚀刻工艺。有关激光蚀刻的进一步讨论，请参见WO 01/25775，其在此全文引入作为参考。最优选地，该技术采用卷带式(reel-to-reel)激光蚀刻工艺。这种工艺可以以卷带方式用来在聚合物基体上形成超薄金属结构，其中金属结构可用作电化学传感器中的电极迹线。采用上述工艺可以将试剂施加到这些结构上。 
令人惊奇的是，已经发现通过采用双面胶带并在电极基体上覆盖部分试剂层，其中所述双面胶带具有各自的切片充当分隔层结构，可以形成传感器的毛细通道和分隔层结构。意想不到的是，在双面胶带覆盖试剂薄膜的位置，没有观察到试样液体的泄漏。所以，能够首先通过激光蚀刻工艺在薄片材料上制备结构化电极迹线，然后模缝涂覆试剂材料，随后通过各自形成的双面胶分隔层限定和血样接触的活性试剂区域。这种工艺可以有利地用来消除生产线的公差。特别地，用分隔层掩盖试剂涂层，可以用来精确限定实际的反应区域。 
在本发明第二方面，本发明涉及采用本发明的剪稀性、轻微触变性或触变性试剂组合物，在固体支撑材料上制备试剂层的方法或工艺。该方法包括提供固体支撑材料，比如塑料材料的薄片，如DuPont的 
Figure S04823924820060303D000071
329。在本发明的工艺中，固体支撑材料相对于模缝涂布头移动。通常，在卷带式工艺中，固体支撑材料横跨模涂布头的狭缝传递。但是，也可以移动模涂布头，而保持薄片材料静止不动。在薄 片材料相对模涂布头的移动过程中，在薄片和模涂布头之间保持限定的距离。优选，该涂覆间距为约30-90μm；通常为68-83μm，最优选为72-76μm。通过迫使试剂组合物通过模缝涂布头的狭缝，将试剂沉积到固体支撑材料上，在固体支撑材料上形成连续的试剂带。如上所述，薄片材料可以包括电极迹线，试剂带可以部分覆盖这些迹线。优选，试剂带在干态下宽度小于1cm，高度小于10μm。 
优选，固体支撑材料以20-80m/min的速度，最优选以30-40m/min的速度相对与模缝涂布头移动。 
优选，试剂组合物以5.5-30g/min，最优选以13-15g/min的涂覆流量输送到固体支撑材料上。 
随后，将沉积的试剂条或者在环境条件下或者在加热的空气流中进行干燥。 
在又一方面，本发明涉及包括上述试剂组合物的分析测试元件。优选，本发明的分析测试元件通过上述工艺制备。 
本发明具有下列优点： 
1、传感器要求的试样体积小(通常100-1000nl)，可以很容易采用模缝涂覆的干薄膜和分隔层/毛细通道叠层工艺构建。该干的薄膜带厚度均匀，而且在整个电化学反应区域均匀。传感器所需的毛细尺寸/不精密度取决于分隔层厚度的变化和毛细通道构造的变化。 
2、模缝涂覆技术可以和电极设计的尖端布线技术配对使用，从而能够在传感器毛细管中实现最小化并获得多重应用，(例如，在合适设计的电极布线中，交错排列两个或多个不同试剂线/条)。可以采用针对两种或多种流体设计的两个交错排列缝模头或者特殊的缝模头组件，实现整个目标。涂覆流体优选具有适当匹配的流变学性质。如果不同流体的涂覆窗口具有一致的重叠区域，那么最佳的技术案例就得以实现。 
3、模缝涂覆薄膜施加技术和试剂的流变学性质配对和组合在一起，使得可以采用卷带式工艺制备用于快速制备诊断用传感器的均匀涂层。 
4、就待涂覆流体的质量分布、其在整个涂覆层上的轮廓、对平整度的影响、可重复性、以及湿层和干层的均匀性而言，所述流体的主要流变学特性是触变或剪稀行为。该特性通过采用黄原胶例如 CMC和二氧化硅，并且浓度和组合与涂覆流体的理想剪稀性、轻微触变性或触变性行为相匹配，而得以实现。 
令人惊奇的是，已经发现二氧化硅，尤其是优选的未处理亲水性二氧化硅，优选颗粒尺寸D50(给，50％的颗粒尺寸是给定尺寸或以下)为1-7μm，在“湿”状态时的任务就是当和薄膜增稠剂 
Figure S04823924820060303D000092
和CMC之一或者全部)组合在一起时，二氧化硅提高了涂覆流体的粘度并改善了涂覆流体的剪稀性、轻微触变性或触变性行为。 
二氧化硅在干态时的作用是，其中： 
a)在如果薄片材料在涂覆和干燥工艺后绕成卷时，防止干薄膜在箔片/载体的未涂覆侧上发生背面传递，和 
b)和光滑涂层相比，增大干涂层的比表面积。不希望受到任何具体理论的束缚，这可能是由于二氧化硅颗粒的颗粒尺寸分布。由于流体传输速度按照该表面积和流体体积之比增加，所以比表面积增大加快了干膜的润湿过程，进而导致毛细填充时间缩短。 
本发明将根据下面的实施例和图进一步阐述。就图而言，任何时候相同的数字、字母和符号代表相同的结构、特征和元件。例如，除非在本申请中有另外说明，下列字码用于： 
1表示薄片； 
2表示溅射的金属薄膜； 
3表示电极对1的工作电极； 
3’表示电极对2的工作电极； 
4表示电极对1的参比/反电极； 
4’表示电极对2的参比/反电极； 
5表示试剂条1； 
5’表示试剂条2； 
6表示分隔层(例如，双面胶) 
7表示顶部箔片；和 
8表示顶部箔片中的通风孔。 
图1和2是在采用本发明的试剂组合物和工艺制备电化学测试元件的工艺中进行的多个步骤的示意图。本领域普通技术人员会很容易认识到本工艺可以用于其它电极构造、具有相同或不同组合物的多个条以及条上的不同位置。应该认识到图1和2所述方法也可以在测试 条上没有电极的情况下执行。本领域普通技术人员会很容易认识到此处所述工艺和试剂也可用于光学测试元件。 
图1和2的部分A和B相同，示出了聚合物薄片(1)，优选是惰性塑料材料，比如Dupont的 
Figure S04823924820060303D000101
329(参见例如，部分A)，在其上面通过常规技术比如溅射、化学、电化学或物理气相沉积等涂覆了金属层(2)(参见例如，部分B)。 
优选，金属层(2)随后通过例如激光蚀刻工艺进行结构化。该工艺去除部分金属层(2)，可以充当电极(3、4)的金属离散结构保留在聚合物薄片(1)的表面上。但是，应该理解的是，也可以采用常规印刷技术或平版印刷工艺在聚合物薄片(1)上创建电极(3、4)。 
在图1的激光蚀刻步骤后，部分C、两个电极(3、4)形成在聚合物薄片上。在图2中，形成的是部分C和两对电极(3、3’、4、4’)。 
在下一步骤中(参见图1和2中的部分D)，在工作电极和反电极(3、3’、4、4’)的活性区域沉积了试剂条5、5’。试剂组合物通过本发明的模缝涂覆工艺施加在电极结构上。 
在图1和2的部分E中，在图1和2的部分D的电极结构上叠层了分隔层(6)。分隔层(6)优选是双面胶带，它覆盖了试剂和电极结构没有和液体试样接触的所有部分。另外，分隔层(6)具有缺口，限定了试剂以及下面电极的反应区域。在分隔层(6)的相反端部，即，电极引线所在的而且没有被试剂组合物覆盖的端部，分隔层(6)留下了电极结构的自由部分，该自由部分可用来将测试条连接到各自的测试条读数计上。 
分隔层(6)优选覆盖试剂(图1中是5，图2中是5’)的窄部分(小于2mm)，从而掩盖试剂涂层的可能的非均匀边缘区域。 
在分隔层(6)叠层到电极和试剂薄片上以后，在优选实施方案中，切除部分薄片材料，对试剂条(5)进行修整。 
在图1和2的部分F中，在分隔层(6)没有和聚合物薄片(1)接触的表面上放置了顶部箔片(7)，优选是惰性塑料盖。聚合物薄片(1)、分隔层(6)和顶部箔片(7)形成了三维毛细通道，该通道由分隔层(6)的厚度以及分隔层切口尺寸限定。为了能够填充毛细空间，优选或者顶部箔片(7)或者聚合物薄片(1)具有通风开孔(8)。 
本领域技术人员会很清楚，聚合物薄片(1)或者顶部箔片(7) 面向毛细空间的表面，可以通过各自的亲水性处理，例如通过涂覆表面活性剂或等离子处理，赋予亲水性。 
实施例 
下面的实施例用于示例而非限制，将更详细公开本发明： 
实施例1：用于电化学安培计葡萄糖生物传感器的试剂组合物。 
制备下列组分的含水混合物： 
Figure S04823924820060303D000111
试剂基质进行定制改性以满足模缝涂覆工艺的要求。向涂层基质中添加二氧化硅、 
Figure S04823924820060303D000112
(黄原胶)、羧甲基纤维素(CMC)和表面活性剂，改变试剂体的流变性。调整表面活性剂浓度，得到位于最优选范围，33-42mN/m的表面张力(用Tensiometer K10T(Kruess)测量)。这个范围的表面张力有助于涂覆条在薄片上的更好粘附以及 受控铺展。采用Rheomat 115(Contraves)测得涂料体的最优选粘度范围是95-115mPa-s。聚合物和二氧化硅也为涂料赋予了触变行为。涂料在通过模缝头分配到薄片上时被剪稀。这样降低了涂料的表观粘度。 
具有这些较低粘度的试剂涂料体的条，在干燥过程中条边缘和试剂组分向着条中心迁移。这种迁移导致在干燥的条中心出现不规则的、不可复制的表面轮廓。具有剪稀性、轻微触变性或触变性质的涂料的分配显示出相同的剪稀效果。但是，涂覆的条在分配以后以及进入干燥区域之前，其粘度立刻回复到接近其表观粘度。在干燥过程中，条边缘向着中心的迁移受到抑制。如图3所示，这导致条的中心、在反应区域出现平整的可重复性区域。更薄的膜进一步抑制了涂覆条的涂层边缘向着中心的迁移。 
实施例2：用于电化学安培计凝结传感器的试剂组合物。 
制备下列组分的含水混合物： 
Figure S04823924820060303D000121
实施例3：用于电化学安培计葡萄糖生物传感器的可替换的试剂组合物。 
制备下列组分的含水混合物： 
实施例4：用于电化学安培计葡萄糖生物传感器的可替换的试剂组合物。 
制备下列组分的含水混合物： 
Figure S04823924820060303D000141
实施例5：涂覆工艺 
聚合物薄片 
Figure S04823924820060303D000142
329，Dupont)在涂覆区域中移动，涂覆区域包括模缝头和支持辊。模缝头(TSE，瑞士)设置零点到薄片表面，经调整后缝和薄片间隙为74μm。为了沉积涂层到薄片上，薄片速度从0上升到38m/min。试剂基质可以采用各种方法包括齿轮泵、活塞、注射器、球囊系统输送到模缝头。调节试剂输送系统，使水流量为13.58ml/min，通过涂布头输送53g/m2的涂料重量。所述涂覆的条的宽度是7.0±0.3mm。涂层在加热的干燥区域中干燥(长度15m、温度110℃、速度为38m/min)，在卷绕点重新卷绕在卷轴上。 
图3-5示出了横跨根据本实施例的试剂条的轮廓曲线测量结果。所用的轮廓测量系统是Dektak IIA Surface Profile Measuring System(Vecco Instruments Inc.，Sloan Technology Division，Dallas，Texas)。Dektak IIA给出的轮廓数据进行了基线校准。 
在图3中，P(mm)代表横跨薄片和试剂条(单位mm)扫描的 x位置，H(μm)代表横跨涂层的各个相对高度(单位μm)。试剂体根据实施例1制备。可以发现，试剂条的横截面宽度约7mm，各个平均中心高度大约5μm。试剂涂层的边缘相对尖锐。涂层的均匀平台区占据了试剂条宽度的大约80％。 
图3所示的试剂涂层的轮廓是本发明涂层的典型轮廓。对于宽度为10mm或更小的试剂条而言，可以获得尖锐的边缘，它在每侧的1mm或以下之内，从下面的薄片材料(对应涂层高度为零)上升到涂层中心的平台区(即，涂层的80％或以上术语均匀的中心平台区)。在中心区域内部，试剂涂层在厚度上实际是均匀的。 
图4给出了横跨根据本实施例制备的试剂条的轮廓曲线测量结果。扫描距离(μm)表示横跨薄片和试剂条扫描的x位置(单位μm)，高度(μm)表示涂层的各自相对高度(单位μm)。试剂体根据采用球磨二氧化硅的实施例1制备。图4A-4E分别给出了涂层重量为20、25、30、40和50mg/m2的结果。 
图5示出了横跨根据对比实施例，即不根据本发明的教导制备的试剂条的轮廓曲线测量结果。扫描距离(μm)表示横跨薄片和试剂条扫描的x位置(单位μm)，高度(μm)表示涂层的各自相对高度(单位μm)。试剂体根据实施例1制备，但没有流变改性剂 CMC和二氧化硅。 
图5A和5B表明，没有流变学改性剂时，干燥的试剂涂层容易在薄片材料上形成不均匀的试剂条。在图5A中，试剂集中在涂覆条的中心部分；在图5B中，试剂集中在位于试剂条的中心和边缘部分之间的两个区域。在两种情况下，边缘部分没有试剂。 
图5(对比实施例)和图3以及4(都根据本发明)的比较结果，显示了本发明的试剂组合物和工艺的有利效果。 
图6是由实施例1(和图4所示的轮廓曲线数据具有可比性)的试剂组合物在薄片材料(中心条周围的亮区)上涂覆的试剂条(中心暗的局限区域)的微观照片。涂覆根据实施例5进行。涂覆的条在横跨涂覆方向(涂覆方向是从顶部到底部)以及沿着涂覆方向都显示出良好的均匀性。 
图7A和7B是涂覆在薄片材料上的、轮廓曲线数据和图5所示的具有可比性的试剂条的微观照片(对比实施例)，和图6所示的光滑 均匀试剂层成强烈对比。涂覆根据实施例5进行；但是，试剂不包含流变学改性剂。涂覆的条在涂覆方向(涂覆方向是从顶部到底部)明显显示出不均匀性。例如，较粘稠的试剂区域由沿着条的暗带表征。图7A显示出在带的中心附近有一条这种暗带(和图5A比较)，而图7B具有两条暗带(和图5B比较)。相信在试剂条内部的该一个或两个较稠试剂涂层区域(暗区)是由于试剂涂层材料的干燥效应。 
实施例6：实施例1的试剂组合物中流变学改性剂的变化。 
在实施例1的试剂组合物中，成分CMC、 
Figure S04823924820060303D000161
Figure S04823924820060303D000162
和PVP的含量根据下表改变。成分含量单位是％w/w，粘度单位是mPa-s。 
Figure S04823924820060303D000163
在本申请中提到的所有公开物、专利和专利申请在此通过引用而结合进来，就好像每份公开物、专利或专利申请都特定而单独指明为全文引入作为参考。 
尽管结合了本发明原理的优选实施方案已经在上面进行了公开，但是本发明不限于这些公开的实施方案。相反，本申请意在覆盖和本公开的下列偏离：在本发明所属的已知或常见现有技术实践之内，而且落在所述权利要求的限制之内。 

Claims (20)

1.一种用于在模缝涂覆操作中制备试剂条的试剂组合物，所述组合物包括试剂、水、改善粘度和触变性的添加剂、和表面活性剂，其中所述组合物是剪稀性或者至少轻微触变性，粘度为70-130mPa.s，而且表面张力为30-50mN/m，其中所述改善粘度和触变性的添加剂选自黄原胶、二氧化硅和羧甲基纤维素。
2.权利要求1的试剂，其粘度为95-115mPa.s。
3.权利要求1或2的试剂，其表面张力为40±2mN/m。
4.权利要求1或2的试剂，其包含二氧化硅。
5.权利要求1或2的试剂，其还包括缓冲剂或者酶或介体或者稳定剂或者蛋白质或者指示剂或者染料或者成膜剂或者辅因子或者所述成分的组合。
6.权利要求1或2的试剂，其包括黄原胶。
7.权利要求1或2的试剂，其包括羧甲基纤维素。
8.权利要求4的试剂，其还包括黄原胶和羧甲基纤维素。
9.在充当固体支撑材料的聚合物薄片上制备试剂层的方法，包括下列步骤：
提供固体支撑材料；
通过移动所述固体支撑材料或者模缝涂布头使所述固体支撑材料相对所述模缝涂布头移动，从而在所述固体支撑材料表面和所述模缝涂布头之间保持限定的距离；
通过所述模缝涂布头沉积试剂组合物，从而在所述固体支撑材料上创建连续的试剂条，其中所述试剂组合物包括试剂、水、改善粘度和触变性的添加剂、和表面活性剂，并且是剪稀性或者至少轻微触变性，粘度为70-130mPa.s，而且表面张力为30-50mN/m，其中所述改善粘度和触变性的添加剂选自黄原胶、二氧化硅和羧甲基纤维素；和
在基片上干燥所述涂覆的试剂条。
10.权利要求9的方法，其中所述试剂条宽度小于1cm。
11.权利要求9的方法，其中所述试剂条高度小于10μm干薄膜厚度。
12.权利要求9的方法，其中所述固体支撑材料相对所述模缝涂布头移动的速度是20-80m/min。
13.权利要求9的方法，其中所述固体支撑材料相对所述模缝涂布头移动的速度是30-40m/min。
14.权利要求9或12的方法，其中所述试剂以5.5-30g/min的涂覆流量输送到所述支撑材料上。
15.权利要求9或12的方法，其中所述试剂以13-15g/min的涂覆流量输送到所述支撑材料上。
16.权利要求9的方法，其中所述模缝涂布头和所述支撑材料之间的距离是30-90μm。
17.分析测试元件，包括：
作为第一固体支撑材料的聚合物薄片；和
在所述第一固体支撑材料上涂覆的试剂条；
特征在于所述试剂条的所述试剂是根据权利要求1-8任一项的试剂。
18.分析测试元件，包括：
作为第一固体支撑材料的聚合物薄片；和
在所述第一固体支撑材料上涂覆的试剂条；
特征在于所述试剂通过根据权利要求9-16任一项的方法涂覆到所述第一固体支撑材料上。
19.权利要求17或18的分析测试元件，其还包括
双面胶带，充当分隔层以限定毛细间隙，而且覆盖部分试剂条；以及
附着到所述分隔层上的第二固体支撑材料，其与所述第一固体支撑材料、所述试剂条以及所述分隔层协作，在试剂条没有被所述双面胶带覆盖的那些部分上创建毛细空间。
20.权利要求17或18的分析测试元件，还包括在所述第一固体支撑材料上的至少两个电极，所述电极部分由所述试剂条覆盖。
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