Author ORCID Identifier
Semester
Summer
Date of Graduation
2025
Document Type
Dissertation
Degree Type
PhD
College
Eberly College of Arts and Sciences
Department
Chemistry
Committee Chair
Peng Li
Committee Member
Stephen Valentine
Committee Member
Glen Jackson
Committee Member
Oluwatobi Odeleye
Committee Member
Kostas Sierros
Abstract
Protein biomarker detection is a critical analytical method for quantifying proteins in biological samples. It is relevant not only in medical diagnostics and research but also numerous other fields such as environmental monitoring, pharmacology, food safety, and biotechnology. Sensitive detection at picogram-per-milliliter (pg/mL) levels is essential for diagnosing diseases, monitoring progression, and tailoring treatments, especially for low-abundance biomarkers like troponin, interleukin 6, carcinoembryonic antigen (CEA), activin A, and vascular endothelial growth factor (VEGF). Immunoassays, particularly enzyme-linked immunosorbent assays (ELISAs), are the gold standard for protein biomarker detection due to their high specificity and sensitivity. The development of point-of-care (POC) diagnostic platforms capable of delivering rapid, accessible, and cost-effective results outside traditional laboratory settings enables timely interventions and expanding diagnostic capabilities to resource-limited areas. The advancement of POC immunoassays relies heavily on innovative device design and surface chemistry to enhance sensitivity, specificity, and usability. The field of 3D printing has been greatly growing over the last decade in terms of device fabrication and application. This fabrication technique offers many advantages over traditional methods that can be taken advantage of to further improve biomarker detection. This work collectively explores the development of 3D-printed microdevices integrated with novel surface modification strategies to improve ELISA for POC applications. In Chapter 2, we establish a surface modification protocol involving air plasma activation followed by GLYMO-labeled streptavidin incubation, which significantly enhances antibody immobilization on various commercial photocurable resins. This strategy improves ELISA performance on both microwells of varying size and microchannels, achieving detection limits comparable to conventional commercial plates. Building on this, Chapter 3 presents a 3D-printed cPlate device coupled with the surface modification method that eliminates the need for magnetic beads, enabling both single biomarker analysis and multiplexed detection within a single device. The immobilization of GLYMO-modified streptavidin and capture antibodies along with the 3D printed fluid manipulation achieved with the cPlate design simplifies ELISA protocols for users and advances the potential for multiplexed POC diagnostics. To address challenges in nonspecific binding and washing steps crucial for POC use, Chapter 4 introduces a liquid-infused surface (LIS) treatment alongside a “key” and well device fabricated through high-resolution LCD printing. This design enables controlled reagent incubation and washing without complex user actions. Functional ELISAs for Activin A detection were optimized on the 3D printed POC ELISA device through the use of the GLYMO surface modification strategy, confirming reproducible, semi-quantitative biomarker detection. Together, these integrated advances in surface chemistry, device architecture, and assay protocols mark significant progress toward practical, sensitive, and user-friendly 3D printed POC ELISA platforms for biomarker detection in healthcare diagnostics. However, in Chapter 5, future directions are discussed on how to further advance this research. One focus would be on the integration of the LID surface with the GLYMO surface modification strategy in the 3D printed POC ELISA device to investigate the capability of going beyond semi-quantitative analysis. Eventually studies will also look into the possibility of adapting the readout methods so that no complex instrumentation is required for the POC setting. Lastly, the potential to turn this into a feasible POC diagnostic test would also then require more investigation of scalability and long-term stability.
Recommended Citation
Binkley, Brandi Marie, "3D Printed Microfluidic Platforms for ELISA-Based Diagnostics: Advancing Toward Point of Care" (2025). Graduate Theses, Dissertations, and Problem Reports. 12931.
https://researchrepository.wvu.edu/etd/12931