Aptamer methods and compositions
Methods of selecting an aptamer that specifically binds to a target molecule complexed with a derivatization agent. Also disclosed are specific aptamers and methods of use thereof.
1. An aptamer selected according to a method comprising:
(i) providing a target molecule;
(ii) providing a derivatization agent;
(iii) contacting the target molecule and the derivatization agent to form a target complex;
(iv) providing an oligonucleotide library comprising a plurality of aptamer candidates;
(v) contacting the target complex and the oligonucleotide library; and
(vi) isolating an aptamer that binds to the target complex; wherein the aptamer does not comprise a cofactor or a derivatization agent.
2. The aptamer of claim 1 , wherein:
the aptamer has a nucleic acid sequence comprising one or more unpaired nucleic acid bases when the aptamer is folded into a double stranded configuration;
the one or more unpaired nucleic acid bases form a binding pocket such that the aptamer can bind a derivatization agent and a target molecule.
3. An aptamer comprising SEQ ID NO: 3; SEQ ID NO: 4 (Glucose-BA_01); SEQ ID NO: 5 (Glucose-BA_07); SEQ ID NO: 6 (Glucose-BA_08); SEQ ID NO: 7 (Glucose-BA_09); SEQ ID NO: 8 (Glucose-BA_10); SEQ ID NO: 9 (Glucose-BA_11); SEQ ID NO: 10 (Glucose-BA_12); SEQ ID NO: 11 (Glucose-BA_13); SEQ ID NO: 12 (Glucose-BA_14); SEQ ID NO: 13 (Glucose-BA_15); SEQ ID NO: 14 (Glucose-BA_16); SEQ ID NO: 15 (Glucose-BA_17); SEQ ID NO: 16 (GLUBA02); SEQ ID NO: 17 (GLUBA09); SEQ ID NO: 18 (GLUBA09_M1); SEQ ID NO: 19 (GLUBA17); SEQ ID NO: 20 (GLUBAN3W10); SEQ ID NO: 21 (GLUBAN3W11); or SEQ ID NO: 22 (GLUBAN3W19), or a sequence at least 90% identical thereto and binding glucose complexed with a bis-boronic derivatization agent.
4. An aptamer comprising SEQ ID NO: 23 (FrucBA02); SEQ ID NO: 24 (FrucBA02_M1); or SEQ ID NO: 25 (FrucBA05), or a sequence at least 90% identical thereto and binding fructose complexed with a bis-boronic derivatization agent.