title: Minimizing antibody cross-reactivity in multiplex detection of biomarkers in paper-based point-of-care assays.
abstract: 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.
authors.affiliation: Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, Sweden. helene.andersson.svahn@scilifelab.se.
authors.forename: J T
authors.initials: JT
authors.lastname: Dias
authors.affiliation: Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, Sweden. helene.andersson.svahn@scilifelab.se.
authors.forename: L
authors.initials: L
authors.lastname: Lama
authors.affiliation: Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, Sweden. helene.andersson.svahn@scilifelab.se.
authors.forename: J
authors.initials: J
authors.lastname: Gantelius
authors.affiliation: Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, Sweden. helene.andersson.svahn@scilifelab.se.
authors.forename: H
authors.initials: H
authors.lastname: Andersson-Svahn
mesh_terms: D055910:Antibodies, Immobilized / Q000276:immunology*; D015415:Biomarkers / Q000097:blood*; D003429:Cross Reactions; D006046:Gold; D006801:Humans; D007118:Immunoassay / Q000379:methods* / Q000706:statistics & numerical data; D057230:Limit of Detection; D053768:Metal Nanoparticles; D019095:Point-of-Care Systems / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D014465:Ultrasonics
chemical_list: D055910:Antibodies, Immobilized; D015415:Biomarkers; D006046:Gold