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Detection of Biomarkers of Periodontal Disease in Human Saliva Using Stabilized, Vertical Flow Immunoassays
ACS Sensors ( IF 8.2 ) Pub Date : 2017-11-13 00:00:00 , DOI: 10.1021/acssensors.7b00745 Emma H. Yee 1 , Shefali Lathwal 1 , Pratik P. Shah 1 , Hadley D. Sikes 1
ACS Sensors ( IF 8.2 ) Pub Date : 2017-11-13 00:00:00 , DOI: 10.1021/acssensors.7b00745 Emma H. Yee 1 , Shefali Lathwal 1 , Pratik P. Shah 1 , Hadley D. Sikes 1
Affiliation
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We report methods for stabilizing cellulose-based immunoassays and using this platform to analyze human saliva. Stabilization treatments of immunoassays for matrix metalloproteinases (MMP)-8 and -9, biomarkers of periodontal disease, were conducted and compared, revealing that anti-MMP-8 and -9 capture antibodies could be stabilized with the addition of a 5% trehalose solution to the test zones, followed by drying in a vacuum oven. After stabilization, the paper devices retained equivalent binding activity to that of freshly prepared tests for 14 days—a time frame that enables US-based clinical testing of this diagnostic assay. A saliva pretreatment method was developed to remove viscous elements without reducing the concentration or binding activity of dissolved proteins. Immunoassays were stored in ziplock bags containing desiccant, and used to detect nanomolar concentrations of MMP-9 in human saliva across the relevant clinical concentration range. These methods and findings facilitate rapid, affordable validation studies of this and other biomarkers that are found in saliva using vertical flow immunoassays.
中文翻译：
稳定的垂直流免疫测定法检测人唾液中牙周疾病的生物标志物
我们报告稳定基于纤维素的免疫分析方法，并使用该平台来分析人类唾液。进行了牙周疾病生物标记物基质金属蛋白酶（MMP）-8和-9免疫测定的稳定化处理并进行了比较，发现加入5％海藻糖溶液可以稳定抗MMP-8和-9捕获抗体到测试区，然后在真空烘箱中干燥。稳定后，纸质设备在14天内保持了与新制备的测试相当的结合活性-这一时间范围使该诊断测定法可在美国进行临床测试。开发了一种唾液预处理方法，以去除粘性成分而不降低溶解蛋白的浓度或结合活性。免疫分析法存放在装有干燥剂的自封袋中，并用于在相关临床浓度范围内检测人唾液中MMP-9的纳摩尔浓度。这些方法和发现促进了使用垂直流免疫测定法对唾液中发现的这种和其他生物标记物进行快速，可负担的验证研究。
更新日期：2017-11-14
中文翻译：
稳定的垂直流免疫测定法检测人唾液中牙周疾病的生物标志物
我们报告稳定基于纤维素的免疫分析方法，并使用该平台来分析人类唾液。进行了牙周疾病生物标记物基质金属蛋白酶（MMP）-8和-9免疫测定的稳定化处理并进行了比较，发现加入5％海藻糖溶液可以稳定抗MMP-8和-9捕获抗体到测试区，然后在真空烘箱中干燥。稳定后，纸质设备在14天内保持了与新制备的测试相当的结合活性-这一时间范围使该诊断测定法可在美国进行临床测试。开发了一种唾液预处理方法，以去除粘性成分而不降低溶解蛋白的浓度或结合活性。免疫分析法存放在装有干燥剂的自封袋中，并用于在相关临床浓度范围内检测人唾液中MMP-9的纳摩尔浓度。这些方法和发现促进了使用垂直流免疫测定法对唾液中发现的这种和其他生物标记物进行快速，可负担的验证研究。
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