Photochemical Sciences Ph.D. Dissertations

First Supramolecular Fluorescence-Based Assay for Carbonic Anhydrase Inhibitors

Date of Award

2016

Document Type

Dissertation

Degree Name

Doctor of Philosophy (Ph.D.)

Department

Photochemical Sciences

First Advisor

Pavel Anzenbacher (Advisor)

Second Advisor

Haowen Xi (Other)

Third Advisor

Marshall Wilson (Committee Member)

Fourth Advisor

Jeremy Klosterman (Committee Member)

Abstract

This work presents supramolecular fluorescent probes capable of specific detection of carbonic anhydrases (CAs), a biologically and clinically important family of metalloenzymes implicated in certain pathological health conditions in humans. The probes, comprising 1,3,4-thiadiazol-5-yl-2-sulfonamide high-affinity moiety, were employed in fluorescence titrations with CAs and showed highly variable analyte-dependent change in fluorescence intensity. Moreover, the probes were utilized in competitive fluorescence-based assay to investigate affinity of model carbonic anhydrase inhibitors towards CAs. The probes were utilized in a competitive sensor array using simple instrumentation under high throughput screening settings. The excellent capability of recognizing a large number of structurally distinct CAIs at various concentrations was demonstrated in qualitative as well as quantitative assays. The throughput, sensitivity and limit of detection surpass the current state-of-the-art methods that generally require enzyme-linked immunosorbent assay based protocols and/or computational modeling. This study opens a new avenue for the development of simple high-throughput assays for the drug development and drugs candidates structure optimization in the near future. To the best of our knowledge, this study presents the first supramolecular fluorescence-based assay for carbonic anhydrase inhibitors.

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