The development of a metalloimmunoassay for the detection of NT-proBNP

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dc.contributor.advisorCrooks, Richard M. (Richard McConnell)
dc.contributor.committeeMemberRichards, Ian
dc.contributor.committeeMemberSchiavinato Eberlin, Livia
dc.contributor.committeeMemberAnslyn, Eric V.
dc.contributor.committeeMemberHoffman, David
dc.creatorPollok, Nicole Elise
dc.date.accessioned2021-07-22T03:16:40Z
dc.date.available2021-07-22T03:16:40Z
dc.date.created2021-05
dc.date.issued2021-05-07
dc.date.submittedMay 2021
dc.date.updated2021-07-22T03:16:40Z
dc.description.abstractThe purpose of this doctoral research is to develop a biosensor for the monitoring of heart failure (HF) in humans. Currently, there is no quantitative patient-facilitated method to monitor HF, and the physical symptoms that result are a poor representation of the acute state of the disease. The biomarker of interest is N-terminal prohormone brain natriuretic peptide (NT-proBNP) which is secreted from the cardiac muscle tissue when the heart is experiencing decompensation. The concentration of NT-proBNP has a direct correlation to the severity of HF, and it is used as the antigen in a metalloimmunoassay, where two monoclonal antibodies are used to sandwich NT-proBNP. One is conjugated to a magnetic microbead via a streptavidin-biotin interaction, and the other is conjugated to a 20 nm-diameter silver nanoparticle (AgNP) using a heterobifunctional cross-linker. The fully formed metalloimmunoassay is placed on a carbon screen-printed and Au electrodeposited sensing electrode to detect AgNP labels electrochemically. Ag charge collected from the assay is representative of the concentration of NT-proBNP in the sample. A phenomenon known as galvanic exchange (GE) is utilized in the detection of Ag. GE is a process that occurs when a zerovalent metal is immersed in a solution containing the oxidized form of a more noble metal. In this specific case, the exchange occurs between AgNP in the metalloimmunoassay and Au³⁺ generated on the sensing electrode. GE occurs because the standard reduction potential of Ag⁺ is slightly lower than Au³⁺. Significant findings of this project reveal that GE between AgNP and Au³⁺ is a process that results in only a partial exchange of AgNP with the Au³⁺ under physiologic conditions. It has also been found that, implementing two subsequent Au³⁺ electrogeneration steps improve the Ag collection efficiency and the reproducibility. Additionally, using heterobifunctional cross-linkers to covalently attach antibodies to AgNP and silver nanocubes (AgNCs) results in a lower limit of detection. These findings have led to the detection of NT-proBNP in buffer within clinically relevant ranges of 0.06-3.49 nM.
dc.description.departmentChemistry
dc.format.mimetypeapplication/pdf
dc.identifier.urihttps://hdl.handle.net/2152/86913
dc.identifier.urihttp://dx.doi.org/10.26153/tsw/13863
dc.language.isoen
dc.subjectElectrochemistry
dc.subjectNT-proBNP
dc.subjectHeart failure
dc.subjectHeterobifunctional cross-linker
dc.subjectPaper-based
dc.subjectSilver nanoparticles
dc.subjectMetalloimmunoassay
dc.subjectGalvanic exchange
dc.titleThe development of a metalloimmunoassay for the detection of NT-proBNP
dc.typeThesis
dc.type.materialtext
thesis.degree.departmentChemistry
thesis.degree.disciplineChemistry
thesis.degree.grantorThe University of Texas at Austin
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy

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