Polymers and conjugates comprising the same
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Publication Number
US-12553886-B2
Publication Date
2026-02-17
Expiration Date
Abstract
Polymers and conjugates comprising polymers are disclosed herein. In some embodiments, the conjugates disclosed are suitable for use as detection probes in immunohistochemical assays, including multiplex immunohistochemical assays.
Core Innovation
The disclosure provides methods for detecting a target in a sample by contacting the sample with a conjugate defined by Formula (VId). The conjugate includes a single-stranded oligonucleotide having specified length and a Tm of less than 70° C., together with a branched or unbranched aliphatic Spacer and a branched or unbranched aliphatic Linker, each optionally having one or more heteroatoms selected from O, N, or S.
The conjugate further includes a Label selected from haptens, fluorophores, chromogens, and enzymes, and a Specific Binding Entity that is an antibody specific for the target. The Oligo, Spacer, or Linker are covalently bonded directly to each other or covalently bonded through an optional group comprising a phosphate group or a phosphodiester group, and detection is performed by detecting the target using the Label.
The framework extends to detecting multiple targets in a sample by contacting the sample with two or more different conjugates, each having Formula (VId) and binding specifically to different targets. Each conjugate comprises different Labels, and the method includes contacting the sample with detection reagents specific to the different Labels, where each detection reagent comprises a different detectable moiety, followed by detecting the multiple targets using the different detectable moieties.
Claims Coverage
The partial content explicitly identifies two independent claims. The main inventive features include a Formula (VId) conjugate with specified oligonucleotide, spacer, linker, and label structure and antibody specificity, and a multiplex detection framework using label-specific detection reagents with different detectable moieties.
Formula (VId) conjugate for target detection with specified oligonucleotide, spacer, linker, and antibody specificity
A method for detecting a target in a sample by contacting the sample with a conjugate having Formula (VId), wherein Oligo is a single-stranded oligonucleotide sequence with specified length and Tm less than 70° C., Spacer and Linker are substituted or unsubstituted branched or unbranched aliphatic groups with specified carbon counts and optional heteroatoms, Label is selected from haptens, fluorophores, chromogens, and enzymes, Specific Binding Entity is an antibody specific for the target, and covalent bonding between Oligo/Spacer/Linker is direct or through an optional phosphate/phosphodiester group; detecting the target using the Label.
Multiplex multiple-target detection using different Formula (VId) conjugates and label-specific detection reagents with different detectable moieties
A method for detecting multiple targets in a sample by contacting the sample with two or more different conjugates each having Formula (VId) that bind specifically to different targets and comprise different Labels; contacting the sample with detection reagents specific to the different Labels where each detection reagent comprises a different detectable moiety; and detecting the multiple targets using the different detectable moieties.
Claim coverage centers on a Formula (VId) conjugate architecture that combines an antibody-specific binding entity with a defined oligonucleotide, spacer, linker, and label arrangement for detecting targets, and a multiplex variant that uses multiple label-specific conjugates together with label-specific detection reagents delivering distinct detectable moieties.
Stated Advantages
Mitigating steric/deactivation effects and background during multiplex detection in formalin-fixed, paraffin-embedded tissue.
Enabling multiplex detection by associating label-specific detection reagents with targets using different detectable moieties.
Documented Applications
Multiplex immunohistochemistry (MIHC) and in situ hybridization (ISH) in formalin-fixed, paraffin-embedded (FFPE) tissue.
Detecting a target in a sample by contacting the sample with a conjugate defined by Formula (VId) and detecting the target using the Label.
Detecting multiple targets in a sample using two or more different conjugates having different Labels and detecting the multiple targets using different detectable moieties.
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