A Method for Designing Instrument-Free Quantitative Immunoassays

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• 作者：Shefali Lathwal ; Hadley D. Sikes
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：March 15, 2016
• 年：2016
• 卷：88
• 期：6
• 页码：3194-3202
• 全文大小：517K
• ISSN：1520-6882

文摘

Colorimetric readouts are widely used in point-of-care diagnostic immunoassays to indicate either the presence or the absence of an analyte. For a variety of reasons, it is more difficult to quantify rather than simply detect an analyte using a colorimetric test. We report a method for designing, with minimal iteration, a quantitative immunoassay that can be interpreted objectively by a simple count of number of spots visible to the unaided eye. We combined a method called polymerization-based amplification (PBA) with a series of microscale features containing a decreasing surface density of capture molecules, and the central focus of the study is understanding how the choice of surface densities impacts performance. Using a model pair of antibodies, we have shown that our design approach does not depend on measurement of equilibrium and kinetic binding parameters and can provide a dynamic working range of 3 orders of magnitude (70 pM to 70 nM) for visual quantification.