![]() 6 This approach gave superior limits of detection but is not suitable for routine, automated, and quantitative analyses in a central clinical laboratory or process analytical technology setting. In our previous work, we improved the sensitivity of an immuno-chromatographic lateral flow assay for MS2 virus detection using functionalized viral nanoparticles as reporters. One particularly robust form of immunoassay is the immunochromatographic lateral flow assay, best-known as the basis of the home pregnancy test. ![]() An alternative approach to the detection of analytes in complex samples is immunoassay, which uses antibodies coupled with highly-detectable reporters to achieve selectivity and sensitivity. Complex clinical and bioprocess samples usually require very high selectivity and substantial sample preparation before chromatographic analysis, 5 however, posing obstacles to the wider use of standard modes of column chromatography. 1– 4 When combined with mass spectrometry, column chromatographic techniques serve as powerful analytical platforms with excellent resolution, sensitivity, and reproducibility. The simple isocratic detection approach described here allows a rapid implementation of immunoassay for detection of a wide range of analytes and uses inexpensive, generally-applicable, and stable column materials instead of costly analyte-specific immunoaffinity adsorbents.Ĭhromatographic analysis of drugs and biomolecules has become increasingly common in the last three decades. This assay principle is demonstrated using M13 bacteriophage virus and human chorionic gonadotropin as model analytes. In the presence of large or artificially-expanded analytes, reporter reagents bind to analyte species to form complexes large enough to be excluded from the adsorbent core, allowing their signal to be observed. In the absence of analyte, antibody-enzyme reporter conjugates can enter the adsorbent and be captured, and their signal is lost. Capto ™ Core 700 and related resins possess a noninteracting size-selective outer layer surrounding a high-capacity nonspecific mixed-mode capture adsorbent core. We introduce analyte-dependent exclusion of reporter reagents from restricted-access adsorbents as the basis of an isocratic reporter-exclusion immunoassay for viruses, proteins, and other analytes.
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