Abstract: It was discovered that hydrogel scaffolds can be used to induce phase separation as aqueous two-phase systems (ATPSs) pass through and/or rehydrate the scaffolds, allowing for concentration of target analyte(s) (e.g., biomolecule(s)) into a particular phase of the ATPS or into a leading front. Accordingly, in various embodiments methods and devices are provided that utilize aqueous two-phase systems and hydrogel scaffolds to improve the sensitivity of assays (e.g., of point-of-care assays) without sacrificing cost or ease of use.

Abstract: In various embodiments single-step ATPS paper-based diagnostic assays are provided that exploit the concept of sequential resolubilization of ATPS components to give rise to the desired phase separation behavior within paper. In one illustrative embodiment, a wick is provided for concentrating an analyte within an aqueous two-phase extraction system in a paper, where the wick comprises a paper configured to receive a sample where the paper comprises a first region containing a first component of an aqueous two-phase system (ATPS) where the first component is in a dry form, and a second region containing a second component of an aqueous two-phase system (ATPS) where the second component is in a dry form; and where said first region and the second region are disposed so that when said wick is contacted with a fluid sample, the first component of said ATPS is hydrated before the second component. In certain embodiments the first and second component are disposed so they are hydrated substantially simultaneously.

Abstract: It was discovered that hydrogel scaffolds can be used to induce phase separation as aqueous two-phase systems (ATPSs) pass through and/or rehydrate the scaffolds, allowing for concentration of target analyte(s) (e.g., biomolecule(s)) into a particular phase of the ATPS or into a leading front. Accordingly, in various embodiments methods and devices are provided that utilize aqueous two-phase systems and hydrogel scaffolds to improve the sensitivity of assays (e.g., of point-of-care assays) without sacrificing cost or ease of use.