A high throughput platform for systematic evolution of ligands by exponential enrichment (SELEX).
The systematic evolution of ligands by exponential enrichment (SELEX) process is a combinatorial chemistry method for the isolation of nucleic acid ligands (aptamers) that bind to a desired target molecule with high affinity. In order to increase throughput via automation, we have adapted the SELEX process for protein targets to a robotics-compatible microtiter plate format. A remarkable feature of the platform is that targets are immobilized by hydrophobic adsorption onto the plate surface. Hydrophobic immobilization procedures are simple and require no specialized modification of the protein target. This format was tested by manually performing four independent SELEX experiments. All were concluded within 8 rounds of selection and yielded aptamers that bind in solution to their respective protein target, calf intestinal alkaline phosphatase, human alpha-thrombin or human platelet derived growth factor, with equilibrium dissociation constants below 3 x 10-10 M.
Drolet
D W
DW
Gilead Sciences, Inc., 2860 Wilderness Place, Boulder, Colorado 80301, USA. ddrolet@nexstar.com
Jenison
R D
RD
Smith
D E
DE
Pratt
D
D
Hicke
B J
BJ
D000469:Alkaline Phosphatase / Q000737:chemistry; D000818:Animals; D002417:Cattle; D020650:Combinatorial Chemistry Techniques; D004268:DNA-Binding Proteins / Q000737:chemistry; D006801:Humans; D008024:Ligands; D009696:Nucleic Acids / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D009843:Oligoribonucleotides / Q000737:chemistry; D010982:Platelet-Derived Growth Factor / Q000737:chemistry; D016601:RNA-Binding Proteins / Q000737:chemistry; D012371:Robotics; D013917:Thrombin / Q000737:chemistry
D004268:DNA-Binding Proteins; D008024:Ligands; D009696:Nucleic Acids; D009838:Oligodeoxyribonucleotides; D009843:Oligoribonucleotides; D010982:Platelet-Derived Growth Factor; D016601:RNA-Binding Proteins; D000469:Alkaline Phosphatase; D013917:Thrombin