Aptamer Selection and Development of Electrochemical Aptamer-based Sensors for Neuropeptide Y

Authors

Evelyn Adela Carreto Guevara, University of Nebraska-LincolnFollow

First Advisor

Rebecca Y. Lai

Committee Members

David Hage, Jiantao Guo

Date of this Version

8-2025

Document Type

Thesis

Citation

A thesis presented to the faculty of the Graduate College at the University of Nebraska in partial fulfillment of requirements for the degree of Master of Science

Major: Chemistry

Under the supervision of Professor Rebecca Y. Lai

Lincoln, Nebraska, August 2025

Comments

Copyright 2025, Evelyn Adela Carreto Guevara. Used by permission

Abstract

In this thesis, we report the results of four aptamers obtained from an aptamer selection process called the systematic evolution of ligands by exponential enrichment (SELEX) with affinity to Neuropeptide Y (NPY). NPY is a highly abundant peptide transmitter with widespread distribution throughout the body which affects various physiological functions. These functions include energy homeostasis, appetite regulation, stress response, and anxiety regulation. Dynamic monitoring of NPY levels is crucial for better understanding behaviors in individuals with neurological disorders associated with NPY. Aptamers are synthetic oligonucleotides obtained through SELEX which have a binding affinity to a target of interest when in their secondary structure. In this thesis, a library consisting of 2 primers and a random domain region of 30 bases for a total of 73 bases was used to identify aptamers for NPY. Upon the completion of 13 SELEX cycles, cloning and sequencing led to 18 individual sequences. Four of these full-length sequences were truncated for analysis against NPY and resulted in dissociation constants between 10 and 60 nM.

Advisor: Rebecca Y. Lai