Polyrotaxanes and uses thereof

Patent metadata

Field	Value
Publication number	US-10118996-B2
Application number	US-201313774302-A
Country	US
Kind code	B2
Filing date	Feb 22, 2013
Priority date	Feb 24, 2012
Publication date	Nov 6, 2018
Grant date	Nov 6, 2018

Abstract

A polyrotaxane containing an affinity binding group has been designed and prepared. The polyrotaxane of the invention can be used for characterization and determination of the three-dimensional structures of biological molecules, such as proteins.

First claim

What is claimed is:

1. A polyrotaxane comprising a plurality of macrocyclic molecules, a linear axle molecule threading through the macrocyclic molecules, and a capping group at each end of the linear axle molecule, wherein one or more of the macrocyclic molecules comprise an affinity binding group and wherein the macrocyclic molecules are laterally and rotationally mobile along the linear axle molecule, wherein the affinity binding group is a metal chelate having a chelating agent moiety and a metal ion, wherein the capping group is linked to the linear moiety through a bond,
wherein said metal ion is Ni 2+ , and
wherein the polyrotaxane has the following structure

wherein x is 113; m is 17; and n is 1.1.

2. A method of characterizing or determining a three-dimensional structure of a biological molecule, comprising the steps of
(a) contacting a polyrotaxane with the biological molecule to form a complex;
(b) subjecting the complex to cryogenic electron microscopy; and
(c) analyzing data collected from the cryogenic electron microscopy experiment wherein said polyrotaxane comprises a plurality of macrocyclic molecules, a linear axle molecule threading through the macrocyclic molecules, and a capping group at each end of the linear axle molecule, wherein one or more of the macrocyclic molecules comprise an affinity binding group and wherein the macrocyclic molecules are laterally and rotationally mobile along the linear axle molecule, wherein the affinity binding group is a metal chelate having a chelating agent moiety and a metal ion, wherein the capping group is linked to the linear moiety through a bond, and
wherein said metal ion is Ni 2+ ;
wherein the polyrotaxane has the following structure

wherein x is 113; m is 17; and n is 1.1.

3. The method of claim 2 , wherein the biological molecule is a protein, a lipid, a nucleic acid, an oligosaccharide, or a complex thereof.

4. The method of claim 2 , wherein the biological molecule is a protein or a complex thereof.

5. The method of claim 2 , wherein the ratio of the number of the biological molecule to the number of the macrocyclic molecules is from about 1:6 to about 1:1.

6. The method of claim 2 , wherein the polyrotaxane is a crosslinked polyrotaxane.

7. The method of claim 6 , wherein the crosslinked polyrotaxane is in the form of a thin membrane for specific capture of affinity tagged biological molecules.

8. The method of claim 7 , wherein the thin membrane is deposited on TEM grids.

Assignees

• Purdue Research Foundation

Inventors

• Thompson David H
• Hyun Seok-Hee
• Wright Kyle J

Classifications

• C08G83/007 Primary

  Polyrotaxanes; Polycatenanes · CPC title

• G01N33/58 Primary

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

• C08B37/0012

  Cyclodextrin [CD], e.g. cycle with 6 units (alpha), with 7 units (beta) and with 8 units (gamma), large-ring cyclodextrin or cycloamylose with 9 units or more; Derivatives thereof · CPC title

• C08B37/0015

  Inclusion compounds, i.e. host-guest compounds, e.g. polyrotaxanes · CPC title

• C08H1/00

  Macromolecular products derived from proteins (food proteins A23; glue, gelatine C09H) · CPC title