Laser-patterned paper-based flow-through filters and lateral flow immunoassays to enable the detection of C-reactive protein
Laser-patterned paper-based flow-through filters and lateral flow immunoassays to enable the detection of C-reactive protein
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DOI：
10.1016/j.talanta.2021.123056
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发表时间：
2021-11-11
期刊：
影响因子：
6.1
通讯作者：
Sones, C. L.
中科院分区：
文献类型：
--
作者：
Galanis, P. P.;Katis, I. N.;Sones, C. L.
我们报告了使用基于激光的制造过程来创建纸基流动过滤器，当与传统的侧向流动免疫分析相结合时，该过滤器提供了一种在较宽浓度范围内检测预定分析物的替代途径。激光图案化的方法被用来创造聚合物结构，改变纸张的孔隙度，以制造具有可控孔隙率水平的多孔流动过滤器。当位于横向流动免疫分析前端的顶端时，流通式过滤器可以阻止超过过滤器有效孔径的颗粒(已知尺寸为200 nm、500 nm、1000 nm和3000 nm)，同时允许较小的颗粒流入横向流动免疫分析。分析物检测基于尺寸专用过滤器的使用，该过滤器保留由目标分析物与两个抗体结合而形成的复合体(大小类似于3µm)，其中每个抗体都标记有不同大小的标记(40 nm Au纳米颗粒和3um m乳胶珠)，并且大于过滤器的有效孔径。该方法用于检测C反应蛋白的浓度范围从10 ng/ml到100,000 ng/ml，检测限发现为13 ng/ml，与其他已报道的用于分析物检测的方法不同，使用该技术我们能够抵消虎克效应，这是许多侧向流动免疫分析中的限制因素。
We report the use of a laser-based fabrication process in the creation of paper-based flow-through filters that when combined with a traditional lateral flow immunoassay provide an alternative pathway for the detection of a pre-determined analyte over a wide concentration range. The laser-patterned approach was used to create polymeric structures that alter the porosity of the paper to produce porous flow-through filters, with controllable levels of porosity. When located on the top of the front end of a lateral flow immunoassay the flow-through filters were shown to block particles (of known sizes of 200 nm, 500 nm, 1000 nm and 3000 nm) that exceed the effective pore size of the filter while allowing smaller particles to flow through onto a lateral flow immunoassay. The analyte detection is based on the use of a size-exclusive filter that retains a complex (similar to 3 mu m in size) formed by the binding of the target analyte with two antibodies each of which is tagged with different-sized labels (40 nm Au-nanoparticles and 3 mu m latex beads), and which is larger than the effective pore size of the filter. This method was tested for the detection of C-reactive protein in a broad concentration range from 10 ng/ml to 100,000 ng/ml with a limit-of-detection found at 13 ng/ml and unlike other reported methods used for analyte detection, with this technique we are able to counter the Hook effect which is a limiting factor in many lateral flow immunoassays.