title: A multiplexed immunoaggregation biomarker assay using a two-stage micro resistive pulse sensor.
abstract: We present an immunoaggregation assay chip for multiplexed biomarkers detection. This chip is based on immunoaggregation of antibody functionalized microparticles (Ab-MPs) to quantify concentrations of multiple biomarkers simultaneously. A mixture of multiple types of Ab-MPs probes with different sizes and magnetic properties, which were functionalized by different antibodies, was used for the multiplexed assay. The interactions between biomarkers and their specific Ab-MPs probes caused the immunoaggregation of Ab-MPs. A two-stage micro resistive pulse sensor was used to differentiate and count the Ab-MP aggregates triggered by different biomarkers via size and magnetic property for multiplexed detection. The volume fraction of each type of Ab-MP aggregates indicates the concentration of the corresponding target biomarker. In our study, we demonstrated multiplexed detection of two model biomarkers (human ferritin and mouse anti-rabbit IgG) in 10% fetal bovine serum, using anti-ferritin Ab and anti-mouse IgG Ab functionalized MPs. We found that the volume fraction of Ab-MP aggregates increased with the increased biomarker concentrations. The detection ranges from 5.2 ng/ml to 208 ng/ml and 3.1 ng/ml to 5.12 × 10(4 )ng/ml were achieved for human ferritin and mouse anti-rabbit IgG. This bioassay chip is able to quantitatively detect multiple biomarkers in a single test without fluorescence or enzymatic labeling process and hence is promising to serve as a useful tool for rapid detection of multiple biomarkers in biomedical research and clinical applications.
authors.affiliation: Department of Mechanical Engineering, University of Akron , Akron, Ohio 44325, USA.
authors.forename: Y
authors.identifier: 0000-0002-1460-7511
authors.initials: Y
authors.lastname: Han
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Akron , Akron, Ohio 44325, USA.
authors.forename: H
authors.initials: H
authors.lastname: Wu
authors.affiliation: Department of Mechanical Engineering, University of Akron , Akron, Ohio 44325, USA.
authors.forename: F
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Akron , Akron, Ohio 44325, USA.
authors.forename: G
authors.initials: G
authors.lastname: Cheng
authors.affiliation: Department of Mechanical Engineering, University of Akron , Akron, Ohio 44325, USA.
authors.forename: J
authors.identifier: 0000-0002-5452-549X
authors.initials: J
authors.lastname: Zhe