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Concept of the “Universal Slope”: Toward Substantially Shorter Decentralized Insulin Immunoassays
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-06-17 , DOI: 10.1021/acs.analchem.2c02178 Eva Vargas 1 , Eleonora M Aiello 2, 3 , Amira Ben Hassine 1 , Victor Ruiz-Valdepeñas Montiel 1 , Jordan E Pinsker 3 , Mei Mei Church 3 , Lori M Laffel 4 , Francis J Doyle 2, 3 , Mary-Elizabeth Patti 4 , Eyal Dassau 2, 3 , Joseph Wang 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-06-17 , DOI: 10.1021/acs.analchem.2c02178 Eva Vargas 1 , Eleonora M Aiello 2, 3 , Amira Ben Hassine 1 , Victor Ruiz-Valdepeñas Montiel 1 , Jordan E Pinsker 3 , Mei Mei Church 3 , Lori M Laffel 4 , Francis J Doyle 2, 3 , Mary-Elizabeth Patti 4 , Eyal Dassau 2, 3 , Joseph Wang 1
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Decentralized sensing of analytes in remote locations is today a reality. However, the number of measurable analytes remains limited, mainly due to the requirement for time-consuming successive standard additions calibration used to address matrix effects and resulting in greatly delayed results, along with more complex and costly operation. This is particularly challenging in commonly used immunoassays of key biomarkers that typically require from 60 to 90 min for quantitation based on two standard additions, hence hindering their implementation for rapid and routine diagnostic applications, such as decentralized point-of-care (POC) insulin testing. In this work we have developed and demonstrated the theoretical framework for establishing a universal slope for direct calibration-free POC insulin immunoassays in serum samples using an electrochemical biosensor (developed originally for extended calibration by standard additions). The universal slope is presented as an averaged slope constant, relying on 68 standard additions-based insulin determinations in human sera. This new quantitative analysis approach offers reliable sample measurement without successive standard additions, leading to a dramatically simplified and faster assay (30 min vs 90 min when using 2 standard additions) and greatly reduced costs, without compromising the analytical performance while significantly reducing the analyses costs. The substantial improvements associated with the new universal slope concept have been demonstrated successfully for calibration-free measurements of serum insulin in 30 samples from individuals with type 1 diabetes using meticulous statistical analysis, supporting the prospects of applying this immunoassay protocol to routine decentralized POC insulin testing.
中文翻译：
“通用斜率”的概念：走向更短的分散式胰岛素免疫测定
如今，在偏远地区对分析物进行分散传感已成为现实。然而，可测量分析物的数量仍然有限，主要是由于需要耗时的连续标准添加校准来解决基质效应并导致结果大大延迟，以及更复杂和更昂贵的操作。这在关键生物标志物的常用免疫测定中尤其具有挑战性，通常需要 60 到 90 分钟才能基于两种标准添加物进行定量，因此阻碍了它们在快速和常规诊断应用中的实施，例如分散式床旁 (POC) 胰岛素测试。在这项工作中，我们已经开发并证明了使用电化学生物传感器（最初开发用于通过标准添加进行扩展校准）在血清样品中为直接免校准 POC 胰岛素免疫测定建立通用斜率的理论框架。通用斜率表示为平均斜率常数，依赖于人血清中 68 种基于标准添加的胰岛素测定。这种新的定量分析方法无需连续添加标准品即可提供可靠的样品测量，从而显着简化和更快的分析（使用 2 个标准添加物时为 30 分钟对比 90 分钟）并大大降低成本，同时不影响分析性能，同时显着降低分析成本.
更新日期：2022-06-17
中文翻译：
“通用斜率”的概念：走向更短的分散式胰岛素免疫测定
如今，在偏远地区对分析物进行分散传感已成为现实。然而，可测量分析物的数量仍然有限，主要是由于需要耗时的连续标准添加校准来解决基质效应并导致结果大大延迟，以及更复杂和更昂贵的操作。这在关键生物标志物的常用免疫测定中尤其具有挑战性，通常需要 60 到 90 分钟才能基于两种标准添加物进行定量，因此阻碍了它们在快速和常规诊断应用中的实施，例如分散式床旁 (POC) 胰岛素测试。在这项工作中，我们已经开发并证明了使用电化学生物传感器（最初开发用于通过标准添加进行扩展校准）在血清样品中为直接免校准 POC 胰岛素免疫测定建立通用斜率的理论框架。通用斜率表示为平均斜率常数，依赖于人血清中 68 种基于标准添加的胰岛素测定。这种新的定量分析方法无需连续添加标准品即可提供可靠的样品测量，从而显着简化和更快的分析（使用 2 个标准添加物时为 30 分钟对比 90 分钟）并大大降低成本，同时不影响分析性能，同时显着降低分析成本.
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