Date of Graduation

5-2020

Document Type

Thesis

Degree Name

Master of Science in Chemical Engineering (MSChE)

Degree Level

Graduate

Department

Chemical Engineering

Advisor/Mentor

Servoss, Shannon L.

Committee Member

Greenlee, Lauren F.

Second Committee Member

Wickramasinghe, S. Ranil

Third Committee Member

Stenken, Julie A.

Keywords

Antigen; Desorption; GFP; Immunosensor; Peptoid; QCM

Abstract

In this work, we have made and characterized a pair of immunobiosensors for detecting the green fluorescent protein (GFP) in an aqueous matrix. An anti-GFP antibody-based biosensor was assembled to detect GFP, while a novel peptoid (N-substituted oligomers of glycine designated as IOS-1) biosensor was also assembled for GFP detection. A quartz crystal microbalance (QCM) gold sensor was used as the supporting substrate for self-assembly of the immunobiosensors. Gravimetric measurements of the QCM gold sensor during immunobiosensor construction and operation were available in real-time using a QCM instrument. X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and Fluorescence microscopy were used to characterize the immunobiosensors. Dose-dependent calibration curves were developed to contrast the performance of the peptoid immunobiosensor and the antibody-based immunobiosensor. The sensitivity of the biosensors shows that the peptoid could detect GFP at 8 nM, unlike the antibody immunobiosensor, which starts to measurably detect GFP at 40 nM. IOS-1 peptoid immunobiosensor had more adsorption capacity for GFP than the antibody-based immunobiosensor and could be reused through multiple adsorption/ desorption cycles. The peptoid immunobiosensor had a binding constant of 2.197 x 10(7) M(-1) with GFP.

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