# US 9,201,067 B2 · Size-controlled macromolecule

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- Number: US 9,201,067 B2
- Document ID: US9201067B2
- Status: Granted
- Kind: B2
- Application number: 10/917,601
- Filed: 2004-08-12
- Published: 2005-02-17
- Granted: 2015-12-01
- Earliest claimed: 2003-09-18
- Assignees: Posco; Postech Foundation
- Applicants: Joon Won Park; Bong Jin Hong; Young Seo Choi; Soon Jin Oh; Kwan Yong Choi
- Inventors: Joon Won Park; Bong Jin Hong; Young Seo Choi; Soon Jin Oh; Kwan Yong Choi
- Web page: https://patentgrep.com/patent/US9201067B2

## CPC

- G01N 33/54353: {with ligand attached to the carrier via a chemical coupling agent (coatings G01N33/54393)}
- C12Q 1/6834: Enzymatic or biochemical coupling of nucleic acids to a solid phase
- 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)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The present application discloses a substrate that includes a molecular layer of regularly spaced size-controlled macromolecules comprising a polymer comprising branched and linear regions in which a plurality of termini on the branched region are bound to the substrate, and a terminus of the linear region is functionalized.

## Claims

### Claim 1

A substrate for detecting the presence of a mutation in a gene, said substrate comprising a molecular layer of a plurality of regularly spaced size-controlled, cone shaped macromolecules, wherein each of said macromolecule comprises a polymer comprising a linear region and a plurality of branched regions in which a plurality of termini on the branched region are covalently bound to the substrate, and the terminus of the linear region is fixed with a target-specific oligonucleotide via a functional group, wherein said polymer is of the formula:
Z—NH—[R1]m—C—{[R2—C]a—{(R3—C)b—[(R4—C)c—(R5—Y)x]y}z}n
wherein
each of m, a, b, and c is independently 0 or 1;
x is 1 when c is 0 or when c is 1, x is 3;
y is 1 when b is 0 or when b is 1, y is 3;
z is 1 when a is 0 or when a is 1, z is 3;
n is 3;
each of R1, R2, R3, R4, and R5 is independently a moiety of the formula
—CH2OCH2CH2C(═O)—;
Z is said target-specific oligonucleotide; and
Y is carboxylic acid terminal group that is bound to the substrate;
provided the product of n, x, y, and z is at least 3, and wherein
said macromolecules are spaced at regular intervals between about 0.1 nm and about 100 nm between the linear functionalized groups.

### Claim 2 (depends on 1)

The substrate according to claim 1, wherein said macromolecules are spaced at regular intervals of about 10 nm.

### Claim 3 (depends on 1)

The substrate according to claim 1, wherein the polymer is a dendron.

### Claim 4 (depends on 1)

The substrate according to claim 1, wherein the substrate is selected from the group consisting of semiconductor, synthetic organic metal, synthetic semiconductor, metal, alloy, plastic, silicon, silicate, glass, and ceramic.

### Claim 5 (depends on 4)

The substrate according to claim 4, wherein the substrate is a slide, particle, bead, micro-well, or porous material.

### Claim 6 (depends on 5)

The substrate according to claim 5, wherein the porous material is a membrane, gelatin or hydrogel.

### Claim 7 (depends on 5)

The substrate according to claim 5, wherein the bead is a controlled pore bead.

### Claim 8 (depends on 1)

A diagnostic system for detecting a mutation in a gene, comprising the substrate according to claim 1.

### Claim 9 (depends on 8)

The diagnostic system according to claim 8, wherein the substrate comprises oligonucleotides that are specific for diagnosis of cancer related genes.

### Claim 10 (depends on 8)

The diagnostic system according to claim 8, wherein the substrate comprises oligonucleotides that are specific for diagnosing mutation in p53 gene.

### Claim 11 (depends on 1)

A method for detecting the presence of a mutation in a gene, comprising contacting a sample containing the gene to be assayed with a substrate according to claim 1, wherein Z is a complementary oligonucleotide of the gene; and determining hybridazation between the gene and the complementary oligonucleotide to detect the presence of a mutation in the gene.

### Claim 12 (depends on 11)

The method according to claim 11, wherein the gene is a cancer related gene.

### Claim 13 (depends on 11)

The method according to claim 11, wherein the gene is p53.