Date of Award

Spring 2018

Degree Name

Master of Arts

Major

Neuroscience

First Advisor

Curran

Second Advisor

Church

Third Advisor

Thilakarathne

Abstract

Polymers imprinted with acetylcholine during synthesis were prepared in order to evaluate their potential for implementation as a novel recognition element in acetylcholine biosensors. Biosensors, such as the glucose monitor, are used to rapidly detect and quantify a target analyte. Acetylcholine biosensors have already been produced using enzymatic recognition elements, but they are currently expensive and plagued by short viability. Molecularly imprinted polymers are not only cheap and durable, but have also been successfully used as a recognition element in biosensors for other analytes. Therefore, computational tools were used to rationally design acetylcholine molecularly imprinted polymers. Three chemicals, itaconic acid, acrylamide, and methacrylamide were identified during this process, because they had an unusually energetically favorable tendency to form a complex with acetylcholine in silico. These three chemicals were used to attempt polymer synthesis in 7mL glass vials, but successful formation was only observed with acrylamide and methacrylamide polymers. A new batch of the two types of polymers was then synthesized and subjected to a binding capacity assay. All polymers were loaded with an 80mM acetylcholine solution, washed three times with deionized water, then washed three times with a designated elution solution. Each sample collected from the polymers during the assay was analyzed via flow injection analysis mass spectrometry. Imprinted polymers generally retained a higher percentage of the acetylcholine they were loaded with than the non-imprinted control polymers. Furthermore, non-imprinted polymers generally had very little acetylcholine left to release after being washed with deionized water, while imprinted polymers still had acetylcholine bound after being washed with deionized water. These results indicated a strong possibility that there was a successful imprinting effect for acetylcholine in the imprinted polymers. A methanol/acetic acid mixture also proved to be the most efficient method for removing acetylcholine from polymers amongst the four elution solutions that were tested. The experimental protocol needs further refinement procedurally and analytically to reliably quantify the acetylcholine in unknown samples from the binding assay. If enough progress is made though, then it could be possible to use the polymers to measure acetylcholine in a solution. This would open up the possibility for acetylcholine molecularly imprinted polymers to be used as an alternative recognition element in acetylcholine biosensors, which have applications in medicine, research, and agriculture.

Comments

Master's Thesis completed at Trinity College, Hartford Connecticut.

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