Vascular endothelial growth factor-binding aptamers

Patent metadata

Field	Value
Publication number	US-9683238-B2
Application number	US-201314655622-A
Country	US
Kind code	B2
Filing date	Dec 12, 2013
Priority date	Dec 28, 2012
Publication date	Jun 20, 2017
Grant date	Jun 20, 2017

Abstract

Disclosed is providing a novel VEGF-binding aptamer whose affinity to VEGF is higher than those of known VEGF-binding aptamers. By an in silico maturation method starting from a known VEGF-binding aptamer, 4 kinds of aptamers whose affinities to VEGF are higher than that of the known VEGF-binding aptamer were prepared. By linking two molecules of an obtained aptamer to each other via a linker, an aptamer having an even higher affinity to VEGF was obtained. The polynucleotide of the present invention contains the base sequence of any one of SEQ ID NOs:1 to 4, and binds to vascular endothelial growth factor.

First claim

The invention claimed is:

1. A polynucleotide comprising the base sequence of any one of SEQ ID NOs:1 to 4, which polynucleotide binds to vascular endothelial growth factor.

2. The polynucleotide according to claim 1 , comprising the base sequence of SEQ ID NO:1.

3. The polynucleotide according to claim 1 , consisting of the base sequence of any one of SEQ ID NOs:1 to 4.

4. The polynucleotide according to claim 3 , consisting of the base sequence of SEQ ID NO:1.

5. The polynucleotide according to claim 1 , wherein a plurality of polynucleotide molecules each consisting of the base sequence of any one of SEQ ID NOs:1 to 4 are linked to each other directly or via a linker(s).

6. The polynucleotide according to claim 5 , wherein two polynucleotides each consisting of the base sequence of SEQ ID NO:1 are linked to each other via a linker.

7. The polynucleotide according to claim 1 , whose dissociation constant for vascular endothelial growth factor is less than 3 nM.

8. The polynucleotide according to claim 7 , whose dissociation constant for vascular endothelial growth factor is less than 1 nM.

9. A method for measuring a concentration of vascular endothelial growth factor in a test sample, said method comprising bringing the polynucleotide according to claim 1 into contact with vascular endothelial growth factor, and measuring binding therebetween.

10. A kit for measuring a concentration of vascular endothelial growth factor, said kit comprising the polynucleotide according to claim 1 .

11. The kit according to claim 10 , comprising a labeled polynucleotide as said polynucleotide.

Assignees

• Nat Univ Corp Tokyo Univ Agriculture & Tech
• Jnc Corp

Inventors

• Ikebukuro Kazunori
• Nonaka Yoshihiko

Classifications

• C12N15/115 Primary

  Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith {; Nucleic acids binding to non-nucleic acids, e.g. aptamers} · CPC title

• C12N2310/16

  Aptamers · CPC title

• G01N33/74

  involving hormones {or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors} · CPC title

• G01N2333/475

  Assays involving growth factors · CPC title

• G01N33/5308

  for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites · CPC title