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Master thesis_SUJI LEE .pdf (2.55 MB)

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Development of pH-sensitive cleavable amine probes for detection of colorectal cancer cell exosome using paper-based immunoassay

Lee, Suji, lee
http://orcid.org/0000-0002-7967-6796
http://rave.ohiolink.edu/etdc/view?acc_num=osu1531993031041248

Abstract Details

2018, Master of Science, Ohio State University, Chemistry.
Due to the emergence of various diseases and the investigation using new therapeutic drugs, immunoassays have been widely used in the biomedical sciences. To achieve certain figures of merit such as; high sensitivity, stability, and selectivity, researchers seek to develop new immunoassay methods. Along with these criteria, early disease detection is important currently which is achieved through point-of-care testing and direct-to-consumer diagnostics. The traditional and commonly used method namely, ELISA (enzyme linked immuno sorbent assay) has several limitations such as the colorimetric based detection, complicated assay process, and limited numbers of available locations for analyzing samples. This thesis focuses on the development of a novel immunoassay platform that is performed on paper substrates and uses pH sensitive cleavable amine probes as mass tags for detection by mass spectrometry. Compared to current traditional method of immunoassay, the proposed platform is expected to emerge as a better alternative by providing a means to achieve high sensitivity, selectivity, rapid analysis, and lower cost. The novelty of the current thesis can be identified as threefold: (1) Two new pH-sensitive amine probes were designed and synthesized for the first time as mass reporter, (2) exosomal HCT116 colorectal cancer cell lines were cultured, isolated, and characterized, and (3) coupled the synthesized amine probes to anti-A33 antibodies and performed immunoassay on paper substrate, after which signal transduction was achieved using mass spectrometry. The probe synthesis was achieved using Steglich esterification process of the selected tertiary amines (N,N-dimethylglycine and 5-(dimethylamino)pentanoic acid) compounds with 4-(2-hydroxyethyl)phenyl isothiocyanate in the presence of 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride (EDC) as a coupling reagent. This yielded iv the incorporation of an ester functional group in the final probe structure, which is sensitive to changes in pHs. Based on the fact that exosomes are detected in most biofluids (blood, urine, or cerebrospinal fluid) and that colorectal cancer is highly ranked among malignant cancers, we believe a low-cost, ultrasensitive test based on exosomal biomarkers will enable early and accurate detection of colorectal cancer, especially among the underserved population (e.g., the uninsured and minorities). Detection of signal from this paper-based immunoassay using mass spectrometry is expected to offer certain advantages such as sensitivity and signal stability over the current immunoassay methods.
Abraham Badu-Tawiah (Advisor)
Robert Baker (Committee Member)
64 p.
Analytical Chemistry

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Citations

  • Lee, lee, S. (2018). Development of pH-sensitive cleavable amine probes for detection of colorectal cancer cell exosome using paper-based immunoassay [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1531993031041248

    APA Style (7th edition)

  • Lee, lee, Suji. Development of pH-sensitive cleavable amine probes for detection of colorectal cancer cell exosome using paper-based immunoassay. 2018. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1531993031041248.

    MLA Style (8th edition)

  • Lee, lee, Suji. "Development of pH-sensitive cleavable amine probes for detection of colorectal cancer cell exosome using paper-based immunoassay." Master's thesis, Ohio State University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=osu1531993031041248

    Chicago Manual of Style (17th edition)

osu1531993031041248
176
© 2018, some rights reserved.Creative Commons License Development of pH-sensitive cleavable amine probes for detection of colorectal cancer cell exosome using paper-based immunoassay by Suji Lee lee is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by The Ohio State University and OhioLINK.