US 12,404,539 B2
Methods for isolating target analytes from urine samples using ATPS and magnetic beads
Cheuk Yiu Tenny Chung, Hong Kong (CN); Vasu Saini, Hong Kong (CN); Daniel William Bradbury, Hong Kong (CN); Harsha Madan Kittur, Singapore (SG); Masae Kobayashi Wen, Buena Park, CA (US); Cheuk Yin Lam, Hong Kong (CN); Kar Kee Tse, Hong Kong (CN); Kit Cheung, Hong Kong (CN); Wing Yee Ng, Hong Kong (CN); Yin To Chiu, Hong Kong (CN); and Garrett Lee Mosley, Hong Kong (CN)
Assigned to PHASE SCIENTIFIC INTERNATIONAL, LTD., Hong Kong (CN)
Filed by Phase Scientific International, Ltd., Hong Kong (CN)
Filed on Sep. 19, 2024, as Appl. No. 18/889,399.
Application 18/889,399 is a division of application No. 18/463,313, filed on Sep. 8, 2023, granted, now 12,129,511.
Claims priority of provisional application 63/381,932, filed on Nov. 2, 2022.
Claims priority of provisional application 63/381,933, filed on Nov. 2, 2022.
Prior Publication US 2025/0011838 A1, Jan. 9, 2025
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6806 (2018.01); C12Q 1/70 (2006.01)
CPC C12Q 1/6806 (2013.01) [C12Q 1/70 (2013.01)] 11 Claims
OG exemplary drawing
 
1. A method for concentrating and purifying at least one target analyte from a clinical biological sample, wherein the clinical biological sample is a urine sample, comprising the steps of
(a) combining the clinical biological sample with a first aqueous two-phase system (ATPS) composition comprising a polymer and a salt component dissolved in an aqueous solution to form a target-rich phase solution and a target-poor phase solution, such that the target analyte is concentrated in the target-rich phase solution, wherein the polymer comprises 5-60% PEG with a molecular weight of at least 400, and the salt component comprises 0.1-6% K2HPO4 and 0.1-18% KH2PO4;
(b) collecting the target-rich phase solution;
(c) adding the target-rich phase solution to a second ATPS composition comprising a second polymer and a second salt component dissolved in an aqueous solution to form a second target-rich phase solution and a second target-poor phase solution, such that the target analyte is concentrated in the second target-rich phase solution, and collecting the second target-rich phase solution, wherein the second polymer comprises 2-25% PEG with a molecular weight of 200 or less, and the second salt component comprises 0.1-50% K2HPO4 and 4-40% KH2PO4;
(d) optionally mixing the target-rich phase solution from step (b) or the second target-rich phase solution from step (c) with a binding buffer to form a mixed solution;
(e) contacting the the second target-rich phase solution from step (c) or the mixed solution from step (d) with a plurality of magnetic beads configured to selectively bind the target analyte such that the plurality of magnetic beads binds to the target analyte; and
(f) eluting and collecting the target analyte from the plurality of magnetic beads with an eluting solution, resulting in a final solution containing the concentrated and purified target analyte.