Minimizing antibody cross-reactivity in multiplex detection of biomarkers in paper-based point-of-care assays.
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Dias JT, Lama L, Gantelius J, Andersson-Svahn H
Nanoscale 8 (15) 8195-8201 [2016-04-21; online 2016-04-01]
Highly multiplexed immunoassays could allow convenient screening of hundreds or thousands of protein biomarkers simultaneously in a clinical sample such as serum or plasma, potentially allowing improved diagnostic accuracy and clinical management of many conditions such as autoimmune disorders, infections, and several cancers. Currently, antibody microarray-based tests are limited in part due to cross reactivity from detection antibody reagents. Here we present a strategy that reduces the cross-reactivity between nanoparticle-bound reporter antibodies through the application of ultrasound energy. By this concept, it was possible to achieve a sensitivity 10(3)-fold (5 pg mL(-1)) lower than when no ultrasound was applied (50 ng mL(-1)) for the simultaneous detection of three different antigens. The detection limits and variability achieved with this technique rival those obtained with other types of multiplex sandwich assays.
PubMed 27030365
DOI 10.1039/c5nr09207h
Crossref 10.1039/c5nr09207h