Department of Chemistry and Biochemistry, North Dakota State University, Fargo, ND 58105, USA; Council for Scientific and Industrial Research- Animal Research Institute, P. O. Box AH20, Achimota, Ghana.
Department of Chemistry and Biochemistry, North Dakota State University, Fargo, ND 58105, USA.
J Pharm Biomed Anal. 2017 Nov 30;146:285-291. doi: 10.1016/j.jpba.2017.09.004. Epub 2017 Sep 5.
A quantitative immunochromatographic assay (QIA) was developed by using gold nanoparticle (GNP)-based lateral flow strip biosensor (LFSB) and a portable strip reader for rapid and sensitive quantitation of Carbohydrate Antigen 19-9 (CA 19-9) in human plasma. CA 19-9 is a biomarker that has been associated with cancers (such as pancreatic and colorectal cancers) and various non-cancerous diseases. The principle is based on sandwich-type immunoreactions between gold nanoparticle (GNP)-labelled detection antibody, anti-CA 19-9 capture antibody and CA 19-9to capture the GNPs on the test zone of LFSB. The accumulation of GNPs on the test zone gave a red line whose intensity was read with a portable strip reader to quantify the concentration of CA 19-9. Assay parameters including the membrane type, antibody concentration, amount of GNP-anti-CA 19-9 conjugates and the components of the running buffer were optimized to obtain the best sensitivity and reproducibility of the assay. The detection limit of the assay was determined to be 5UmL (S/N=3) with a linear range of 5UmL-100UmL. CA 19-9 concentrations in healthy human and pancreatic cancer patient plasma samples were successfully evaluated using the developed quantitative immunochromatographic assay (QIA), and the results were in accordance with that obtained with enzyme linked immunosorbent assay (ELISA). The developed assay shows great promise for clinical application and biomedical diagnosis, particularly in limited resource settings.
一种基于金纳米粒子（GNP）的横向流条生物传感器（LFSB）和便携式条带读取器的定量免疫层析分析（QIA）已被开发出来，用于快速灵敏地定量检测人血浆中的碳水化合物抗原 19-9（CA 19-9）。CA 19-9 是一种与癌症（如胰腺癌和结直肠癌）和各种非癌症疾病相关的生物标志物。其原理基于金纳米粒子（GNP）标记的检测抗体、抗 CA 19-9 捕获抗体和 CA 19-9 之间的三明治型免疫反应，以捕获 LFSB 测试区的 GNP。测试区 GNP 的积累产生了一条红线，其强度用便携式条带读取器读取，以定量 CA 19-9 的浓度。优化了膜类型、抗体浓度、GNP-抗 CA 19-9 缀合物的量以及运行缓冲液的成分等测定参数，以获得该测定法的最佳灵敏度和重现性。该测定法的检测限为 5UmL（S/N=3），线性范围为 5UmL-100UmL。使用开发的定量免疫层析分析（QIA）成功评估了健康人及胰腺癌患者血浆样本中的 CA 19-9 浓度，其结果与酶联免疫吸附测定（ELISA）获得的结果一致。该开发的测定法在临床应用和生物医学诊断方面具有广阔的应用前景，特别是在资源有限的环境中。
