Aptamer selection method
AssigneeMcMaster University
InventorsYingfu Li, Razvan Nutiu
Life of this patent
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Abstract
The present invention provides a method for the selection of aptamer sequences and aptamers identified using the method. The method comprises immobilizing a library oligonucleotide by forming a duplex with an antisense oligonucleotide, reating the duplex molecule with a target and collecting oligonucleotide molecules which dissociate from the duplex structure.
Claims16
A method for selecting aptamer capable of binding to a target, said method comprising the steps of:
- i) interacting antisense oligonucleotide having the nucleotide sequence of SEQ ID NO:
- 1 and being adapted to be attached to a solid support with an oligonucleotide library containing multiple oligonucleotides, each of which includes an antisense binding domain having a sequence complementary to SEQ ID NO:
- 1 and a random nucleotide domain, wherein the antisense oligonucleotide and the oligonucleotides in the library form double stranded duplexes, ii) immobilizing the duplexes on a solid support;
- iii) incubating the duplexes in the presence of the target;
- and iv) collecting library oligonucleotides that dissociate from the duplexes and bind to the target.
A method for the selection of an aptamer specific for a target, said method comprising the steps of:
- i) providing an oligonucleotide library containing multiple oligonucleotides, each of which includes an antisense binding domain having a sequence complementary to SEQ ID NO:
- 1, at least one random sequence domain, a 3′ primer binding domain and a 5′ primer binding domain;
- ii) hybridizing the oligonucleotides in the library to a biotinylated antisense oligonucleotide having the nucleotide sequence of SEQ ID NO:1 to form duplex molecules on beads;
- iii) incubating the beads of ii) with the target;
- iv) collecting oligonucleotides which have bound to the target;
- and v) amplifying the collected oligonucleotides.
An aptamer selection system comprising an antisense oligonucleotide having the nucleotide sequence of SEQ ID NO:1, and an oligonucleotide library containing multiple oligonucleotides, each of which includes an antisense binding domain having a sequence complementary to SEQ ID NO:1 and at least one random nucleotide domain, wherein said antisense oligonucleotide is adapted to be attached to a solid support.
An aptamer selection system comprising an antisense oligonucleotide comprising SEQ ID. NO. 1, an oligonucleotide library containing multiple oligonucleotides each of which includes SEQ ID. NO. 2, a P1 oligonucleotide comprising SEQ ID. NO. 3, a P2 oligonucleotide comprising SEQ ID NO. NO. 4, and a P3 oligonucleotide comprising SEQ ID. NO. 5.
Dependencies are inferred from the claim text.