Arrastia Mary, Avoundjian Ani, Ehrlich Paul Said, Eropkin Micah, Levine Leanna, Gomez Frank A
Department of Chemistry and Biochemistry, California State University, Los Angeles, CA, USA.
Electrophoresis. 2015 Mar;36(6):884-8. doi: 10.1002/elps.201400421. Epub 2015 Feb 20.
A novel microfluidic paper-based analytical device (μPAD) utilizing a nitrocellulose (NC) membrane to detect IgG antibodies through a colorimetric analysis is described. The μPAD was constructed using layered polyethylene terephthalate (PET) and pressure-sensitive adhesives (PSA). The biotin labeled Goat Anti-Mouse IgG antibody was spotted and dried on the NC channel prior to subjecting it to a series of wash solutions (Tris-tween), increasing concentrations of alkaline phosphatase conjugated to streptavidin (Strep-ALP), and para-nitrophenyl phosphate (p-NPP) realizing a vibrant yellow color. The reaction proceeds for 10 min before applying the p-NPP stop solution. The device was then dried, scanned, and analyzed yielding a linear range of inverse yellow color intensities versus Strep-ALP concentrations. The development of this simple μPAD should further facilitate the use of NC in colorimetric assays to detect and quantitate antibodies.
描述了一种新型的基于微流控纸的分析装置（μPAD），该装置利用硝酸纤维素（NC）膜通过比色分析来检测IgG抗体。μPAD是使用层状聚对苯二甲酸乙二醇酯（PET）和压敏粘合剂（PSA）构建的。在将生物素标记的山羊抗小鼠IgG抗体置于一系列洗涤溶液（Tris-吐温）、浓度不断增加的与链霉亲和素偶联的碱性磷酸酶（Strep-ALP）以及对硝基苯磷酸酯（p-NPP）之前，先将其点样并干燥在NC通道上，从而实现鲜黄色。在加入p-NPP终止溶液之前，反应进行10分钟。然后将装置干燥、扫描并分析，得出黄色强度倒数与Strep-ALP浓度之间的线性范围。这种简单的μPAD的开发应进一步促进NC在比色测定中用于检测和定量抗体。
