Image differentiated multiplex assays
Inventors
Tsao, Dean • Huang, Chin-Shiou • CHUNG, Yao-Kuang
Assignees
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Publication Number
US-11579522-B2
Publication Date
2023-02-14
Expiration Date
Abstract
Provided herein are encoded microcarriers for analyte detection in multiplex assays. The microcarriers are encoded with an analog code for identification and include a capture agent for analyte detection. Also provided are methods of making the encoded microcarriers disclosed herein. Further provided are methods and kits for conducting a multiplex assay using the microcarriers described herein.
Core Innovation
The invention relates to an encoded microcarrier built from a substantially transparent polymer layer with a first surface and a second surface that are parallel to each other. A discontinuous, substantially non-transparent polymer layer is affixed to the first surface of the transparent polymer layer so that the non-transparent layer encloses a center portion of the substantially transparent polymer layer. The discontinuous substantially non-transparent polymer layer represents an analog code for analog identification via analog code carried by the microcarrier.
A capture agent is coupled to at least one of the first surface and the second surface of the substantially transparent polymer layer in at least the center portion of the substantially transparent polymer layer. The capture agent is configured for capturing an analyte, with the coupling occurring in the coded center portion of the transparent polymer layer. The coded center portion and coupled capture agent together define the functional association between the analog code and analyte binding.
The platform further includes multiplex detection concepts using encoded microcarriers with analog codes, including decoding codes and detecting bound analytes with fluorescence microscopy and luminescence microscopy options as described. The non-transparent analog code is optionally combined with magnetic and nickel layer concepts to support orientation and magnetic separation steps, together with optional projecting columns to prevent sticking. In example microcarrier designs, the non-transparent analog code can be implemented using a gear-shaped two-dimensional analog encoding in which the analog code is specified by gear-teeth dimensions and arrangement.
Claims Coverage
The document provides two independent claims that cover (i) a single encoded microcarrier and (ii) a plurality of encoded microcarriers. Across these independent claims, there are three core inventive features recurring in each: a substantially transparent polymer layer, a discontinuous substantially non-transparent polymer layer that represents an analog code and encloses a center portion, and a capture agent coupled to the transparent polymer in at least the center portion to capture an analyte.
Substantially transparent polymer layer with parallel first and second surfaces
A substantially transparent polymer layer having a first surface and a second surface, the first and the second surfaces being parallel to each other.
Discontinuous substantially non-transparent polymer layer representing an analog code enclosing a center portion
A substantially non-transparent polymer layer, wherein the substantially non-transparent polymer layer is a discontinuous, substantially non-transparent polymer layer affixed to the first surface of the substantially transparent polymer layer that encloses a center portion of the substantially transparent polymer layer, wherein the discontinuous, substantially non-transparent polymer layer represents an analog code.
Capture agent coupled to the transparent polymer in the center portion for analyte capture
A capture agent for capturing an analyte, wherein the capture agent is coupled to at least one of the first surface and the second surface of the substantially transparent polymer layer in at least the center portion of the substantially transparent polymer layer.
Plurality of encoded microcarriers with analyte-capturing coded microcarriers
A plurality of encoded microcarriers, wherein each encoded microcarrier comprises: (a) a substantially transparent polymer layer having a first surface and a second surface, the first and the second surfaces being parallel to each other; (b) a substantially non-transparent polymer layer affixed to the first surface enclosing a center portion of the substantially transparent polymer layer, wherein the discontinuous substantially non-transparent polymer layer represents an analog code; and (c) a capture agent for capturing an analyte, coupled to at least one of the first surface and the second surface of the substantially transparent polymer layer in at least the center portion of the substantially transparent polymer layer.
Overall claim coverage centers on encoded microcarriers having a substantially transparent polymer layer with parallel surfaces, a discontinuous substantially non-transparent polymer region that encloses a center portion and represents an analog code, and a capture agent coupled to the transparent polymer in at least the center portion to capture an analyte. The plurality claim extends the same microcarrier structure to multiple encoded microcarriers, and dependent claims further distinguish microcarrier species by different analog codes and different corresponding capture agents and analytes.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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