A three arm y-shaped bisbiotin ligand

Patent metadata

Field	Value
Publication number	US-2017067902-A1
Application number	US-201515119549-A
Country	US
Kind code	A1
Filing date	Feb 18, 2015
Priority date	Feb 18, 2014
Publication date	Mar 9, 2017
Grant date	—

Abstract

Embodiments of the present disclosure provide bisbiotin ligands and related conjugates and methods. The bisbiotin ligands, combined with streptavidin, can be used in the separation, labelling, targeting, and immobilization of biomolecules.

First claim

1 . A ligand comprising a multiplicity of biotin moieties, wherein each biotin moiety is attached to a rigid, nano-meter central core through a flexible linker, wherein the ligand presents at least two biotin moieties to a target.

2 . The ligand of claim 1 , wherein the target is streptavidin.

3 . The ligand of claim 1 , wherein two biotin moieties are attached to a central trilinker.

4 . The ligand of claim 3 , wherein the trilinker comprises the structure

5 . The ligand of claim 3 , wherein the trilinker comprises the structure

6 . The ligand of claim 3 , wherein the trilinker comprises the structure

7 . The ligand of claim 4 , 5 or 6 , wherein n is 1 to 50.

8 . The ligand of claim 3 , wherein the ligand comprises the structure

9 . The ligand of claim 8 , wherein n is 1 to 50.

10 . The ligand of claim 3 , wherein the ligand is conjugated to a moiety selected from the group consisting of: fluorescent tags, quantum dots, redoxes, magnetic nanoparticles, and gold nanoparticles.

11 . The ligand of claim of claim 10 , wherein the ligand comprises the structure

12 . The ligand of claim 1 , wherein the ligand is immobilized to an atomic force microscopy tip.

13 . The ligand of claim 12 , wherein the ligand is connected to the atomic force microscopy tip via a PEG linker

14 . The ligand of claim 1 , wherein the ligand is attached to nanoparticles functionalized with alkynes.

15 . A method for the separation and detection of biomolecules, the method comprising:
immobilizing streptavidin to the ligand of claim 14 ; attaching one or more biotinylated probes or affinity molecules to form a streptavidin/biotin nanoparticle complex; incubating the nanoparticle complex with a biological sample under conditions to allow the nanoparticle complex to bind to biomolecules in a biological sample; detecting the binding of biomolecules to the nanoparticle complex; and separating the biomolecules bound to the nanoparticle complex from the biological sample.

16 . The method of claim 15 , wherein the biomolecule is selected from the group consisting of DNA, RNA, proteins, and peptides.

17 . The method of claim 15 , wherein the probes are DNA probes or RNA probes.

18 . The method of claim 15 , wherein the affinity molecules arc one or more of antibodies, aptamers, or peptides.

19 . The method of claim 15 , wherein the biotinylated DNA probes or affinity molecules are conjugated with ligand of claim 1 .

20 . The ligand of claim 1 , further comprising a thiol group at a non-biotin moiety region of the ligand.

21 . The ligand of claim 20 , wherein the ligand comprises the structure

22 . A monolayer comprising a gold substrate, wherein the ligand of claim 20 is immobilized thereto.

23 . The monolayer of claim 22 , wherein the monolayer further comprises an alkylated PEG spacer.

Assignees

• Univ Arizona State

Inventors

• Zhang Peiming
• Lindsay Stuart
• Senapati Subhadip
• Biswas Sudipta

Classifications

• C07C247/04

  being saturated · CPC title

• G01N33/58

  involving labelled substances (G01N33/53 takes precedence) · CPC title

• G01Q60/38

  Probes, their manufacture, or their related instrumentation, e.g. holders · CPC title

• C12Q1/6816

  characterised by the detection means (C12Q1/6804 takes precedence) · CPC title

• G01N33/587 Primary

  Nanoparticles · CPC title