Developing precise and accurate lateral flow assays (LFAs) requires the systematic optimization of multiple critical components. This process involves selecting the appropriate antibody pair, conjugation conditions, sample and conjugate pad materials, nitrocellulose membrane, test line concentration, wick pad material, blocking reagents and buffers, cassette, and sample volume. Additionally, for quantitative assays, the standardization analyte and matrix must also be optimized. These components are interrelated and require careful balance to produce effective and functional assays.
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