# US 10,118,996 B2 · Polyrotaxanes and uses thereof

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- Number: US 10,118,996 B2
- Document ID: US10118996B2
- Status: Granted
- Kind: B2
- Application number: 13/774,302
- Filed: 2013-02-22
- Published: 2013-08-29
- Granted: 2018-11-06
- Earliest claimed: 2012-02-24
- Assignees: Purdue Research Foundation
- Applicants: Purdue Research Foundation
- Inventors: David H. Thompson; Seok-Hee Hyun; Kyle J. Wright
- Web page: https://patentgrep.com/patent/US10118996B2

## CPC

- C08G 83/007: {Polyrotaxanes; Polycatenanes}
- C08B 37/0012: {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}
- C08B 37/0015: {Inclusion compounds, i.e. host-guest compounds, e.g. polyrotaxanes}
- C08H 1/00: Macromolecular products derived from proteins (food proteins A23; glue, gelatine C09H)
- G01N 33/58: involving labelled substances (G01N33/53 takes precedence)
- C08G: MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08B: POLYSACCHARIDES; DERIVATIVES THEREOF (polysaccharides containing less than six saccharide radicals attached to each other by glycosidic linkages C07H; fermentation or enzyme-using processes C12P19/00; sugar industry C13; production of cellulose D21)
- C08H: DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS (polysaccharides C08B; natural rubber C08C; natural resins or their derivatives C09F; bituminous materials C10)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## 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.

## Claims

### Claim 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 Ni2+, and
wherein the polyrotaxane has the following structure
wherein x is 113; m is 17; and n is 1.1.

### Claim 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 Ni2+;
wherein the polyrotaxane has the following structure
wherein x is 113; m is 17; and n is 1.1.

### Claim 3 (depends on 2)

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

### Claim 4 (depends on 2)

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

### Claim 5 (depends on 2)

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.

### Claim 6 (depends on 2)

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

### Claim 7 (depends on 6)

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.

### Claim 8 (depends on 7)

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