Ultra-sensitive analyte detection and quantification using catch and release with proximity detection
AssigneeChildren's Medical Center Corporation
InventorsClinton H. Hansen, Wesley Philip Wong
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Abstract
Provided herein is a highly sensitive analyte detection method that involves catch and release of a target analyte coupled with a proximity assay. Target analyte is captured using a capture binding partner and is also bound by detection binding partners thereby forming an analyte-biding partner complex. The complex is immobilized, and unbound reactants are physically removed. Thereafter the complex is released and a proximity assay is performed on the complex in order to detects its presence.
Claims23
A method of detecting a target analyte, comprising
- combining a capture binding partner and a detection binding partner with a sample, under conditions sufficient for the capture binding partner and detection binding partner to bind to a target analyte present in the sample, thereby forming an analyte-binding partner complex
- immobilizing the analyte-binding partner complex to a solid support,
- removing unbound capture binding partner and detection binding partner,
- releasing the analyte-binding partner complex,
- transferring the released analyte-binding partner complex to a second container
- performing a proximity assay on the released analyte-binding partner complex in solution, thereby detecting the presence of the analyte-binding partner complex.
A method of detecting a target analyte, comprising
- forming an analyte-binding partner complex comprising a target analyte bound simultaneously to a capture binding partner and two or more detection binding partners, wherein the capture binding partner is immobilized to a solid support before or after binding to the target analyte,
- separating the analyte-binding partner complex from unbound capture binding partner and/or unbound detection binding partner,
- releasing immobilized analyte-binding partner complex,
- transferring the released analyte-binding partner complex to a second container
- performing a surface-free proximity assay on the released analyte-binding partner complex, thereby detecting the presence of the analyte-binding partner complex.
Dependencies are inferred from the claim text.