US 12,258,558 B2
Compositions and methods for screening aptamers
Trevor Feagin, Stanford, CA (US); Diana Wu, Stanford, CA (US); Peter Mage, Stanford, CA (US); John Coller, Stanford, CA (US); and Hyongsok Tom Soh, San Francisco, CA (US)
Assigned to CZ Biohub SF, LLC, San Francisco, CA (US); and The Board of Trustees of The Leland Stanford Junior University, Stanford, CA (US)
Appl. No. 15/734,512
Filed by CZ Biohub SF LLC, San Francisco, CA (US); and The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
PCT Filed Jun. 4, 2019, PCT No. PCT/US2019/035345
§ 371(c)(1), (2) Date Dec. 2, 2020,
PCT Pub. No. WO2019/236548, PCT Pub. Date Dec. 12, 2019.
Claims priority of provisional application 62/680,435, filed on Jun. 4, 2018.
Prior Publication US 2021/0324374 A1, Oct. 21, 2021
Int. Cl. C12N 15/10 (2006.01); C40B 30/04 (2006.01)
CPC C12N 15/1048 (2013.01) [C40B 30/04 (2013.01)] 24 Claims
OG exemplary drawing
 
1. A method for screening a library of aptamers for aptamers having binding affinity to a target molecule, comprising:
(a) providing a solid support comprising clusters of sequence-identified adaptor end- linked aptamers,
wherein each cluster comprises a plurality of identical sequence-identified adaptor end-linked aptamers comprising one or more non-natural nucleotide covalently bonded to a binding agent, and optionally extended solid support-linked oligonucleotides comprising complements of the adaptor end-linked aptamers, and wherein the sequence of the adaptor end-linked aptamers in each cluster is unique compared to that of the other clusters;
(d) probing the clusters of adaptor end-linked aptamers linked to the solid support with a target molecule;
(e) washing unbound target molecule from the solid support;
(f) screening the solid support for binding and location of the target molecule; and
(g) correlating the location of binding of the target molecule to the nucleotide sequence of an solid support-linked aptamer at the location of binding of the target molecule, thereby identifying the nucleotide sequence of an aptamer that binds the target molecule.