Time Stability of the Binding Affinity of 5-Hydroxytryptophan to DFHBI- 1T- Broccoli RNA Complex Using Fluorescence Detection
Neville, Alaya M.
Aptamers are an alternative to antibodies with a longer shelf life, multi-diverse target capabilities, effective production rate, and thermal flexibility. Our focus was to explore the optimal conditions and methods for our DFHBI-1T RNA biosensors to bind to the serotonin precursor 5-hydroxytryptophan (5-HTP) in vitro that can mimic in vivo conditions. We tracked our fluorescent markers and determined their binding affinities via fluorescence-based assays. Through a series of 15-minute interval timed assays set at standard conditions we found that when measured at 25Æ C, the binding affinity of biosensors CM 2, CM 5, CM 6, CM 7 and CM 8 did not change over a 45- minute time period, while biosensors CM 3 and CM 4 did change. Conversely, we found that when measured at 37Æ C, biosensors CM 3, CM 4 and CM 5 change over a 45- minute period; CM 2 and CM 5- CM 8 did not.
vii, 20 p.
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