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This study introduces a novel "graded ladder bar" format for lateral flow assays (LFAs), enabling accurate quantification of analyte concentrations. By tuning test line sensitivity, LFAs can now provide quantitative results over a wide dynamic range.

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Area of Science:

  • Biomolecular assays
  • Analytical chemistry
  • Biosensor technology

Background:

  • Lateral flow assays (LFAs) are common for qualitative yes/no diagnostic tests.
  • Quantification using LFAs is challenging due to limitations in sensitivity and dynamic range.

Purpose of the Study:

  • To develop a tunable lateral flow assay (LFA) for accurate quantification of sample concentrations.
  • To introduce a novel "graded ladder bar" format for enhanced LFA performance.

Main Methods:

  • Tuning test line sensitivity by varying capture molecule density.
  • Developing a mass-action kinetics model to predict assay performance.
  • Designing and testing a three-line "graded ladder bar" strip with logarithmically spaced concentration thresholds.

Main Results:

  • Test line sensitivity in LFAs can be precisely tuned by adjusting capture molecule density.
  • A kinetic model accurately predicted LFA signal and shape across various conditions.
  • The "graded ladder bar" format demonstrated accurate, logarithmically spaced detection thresholds, matching model predictions.

Conclusions:

  • The "graded ladder bar" format enables quantitative detection in LFAs by controlling capture efficiency.
  • This approach offers a robust method for designing nonlinear responses and expanding the dynamic range of LFAs.
  • The developed format is a valuable tool for applications requiring precise quantification of sample concentrations.