Design, synthesis and use of synthetic nucleotides comprising charge mass tags

Inventors

O'Halloran, Jonathan

Assignees

QuantumDx Group Ltd

Abstract

Embodiments of the present disclosure relate generally to reporter compositions which are synthetic nucleotides that comprise nucleotides with a high charge mass moiety attached thereto via a linker molecule. The linker molecules can vary in length in part to enable the high charge mass moiety to extend out from a DNA polymerase complex so that polymerization may not be influenced.

Core Innovation

The invention provides a reporter composition that includes a nucleotide or its derivative, a charged moiety, and a linker molecule attached to the nucleotide or its derivative and the charged moiety. The charged moiety comprises an electrical charge sufficient to change an electrical property of a sensitive detection nanostructure when the reporter composition is operably coupled to the sensitive detection nanostructure.

In specific embodiments, the linker molecule uses an H2N–L–NH2 diamine framework, where L is a linear or branched chain that includes an alkyl group and/or an oxy alkyl group. The charged moiety comprises one or more groups, derivatives, and combinations thereof that enable the electrical-property change, including architectures comprising an aromatic and/or aliphatic skeleton with tertiary amino groups, alcohol hydroxyl groups, and phenolic hydroxy groups.

Additional embodiments describe reporter compositions in which the charged moiety has pH-dependent electrical charge behavior and is configured not to affect nucleotide polymerization by a polymerase. The disclosed concept is also tied to sequencing by synthesis using charge detection, where cleavable-linker or removable reporter moiety behavior enables read-out based on electrical changes near the sensitive nanostructure surface.

Claims Coverage

The provided independent claims include reporter-composition claim families and an additional independent claim that limits the reporter moiety R selection and ties the reporter composition to a kit for determining a nucleotide sequence. Across these, the inventive features center on the reporter composition structure, the charged moiety electrical charge sufficient to change a sensitive detection nanostructure property, the linker and charged-moiety group composition, optional pH-variable charge and non-interference with polymerase polymerization, and the allowed reporter moiety set.

The claim coverage centers on reporter compositions that physically link a nucleotide or derivative to a charged moiety via a linker, where the charged moiety’s electrical charge is sufficient to change the electrical property of a sensitive detection nanostructure. Additional claim features narrow the linker to an H2N–L–NH2 diamine framework with defined alkyl/oxy-alkyl chain content, define charged-moiety group composition and pH-variable charge behavior, and require the charged moiety not to affect nucleotide polymerization by a polymerase. Separate independent claims broadly limit the reporter component R to a defined set of nucleotide-like entities and, for one family, embed the reporter composition in a kit for determining a nucleotide sequence.

Stated Advantages

Documented Applications