Routine high-sensitivity multiplexed detection of disease biomarkers in blood will have an impact on early diagnosis and therapy selection. A novel and highly effective approach for detection of broad categories of disease is multiplexed detection of antibodies using protein microarrays. As a greater number of antibody biomarkers are being identified and clinically validated for viral infection, cancer, cardiovascular disease, and autoimmune disease, there is a strong need to detect them with greater sensitivity and greater signal-to-noise ratio, while at the same time being able to perform automated biomarker analysis with low cost per test. In the proposed project, the investigators integrate photonic crystal enhanced fluorescence (PCEF) technology with a novel size-exclusion blood filtration technology to develop a multiplexed microspot fluorescent sandwich assay platform for the clinical laboratory environment. Our approach integrates several innovations into an inexpensive plastic-based sensor cartridge and desktop detection instrument: 1. A silicon-based nanostructured resonant optical photonic crystal chip with an integrated Fabry-Perot optical cavity will be used to deliver ~50x greater signal enhancement than current-generation PCEF devices, which already routinely provide <1 pg/ml limits of detection in complex media, using only 10 ?l sample volumes. 2. A laser-machined blood filter will separate plasma from a droplet of heparinized whole blood in 60 seconds, enabling the entire assay protocol to be performed automatically in <60 minutes without user intervention. 3. An innovative laser scanning approach is used to couple light from a small semiconductor laser directed to the PC surface in the optimal ?on-resonance? condition, resulting in a rugged, compact instrument with a cost of <$10K. 4. A statistical bioinformatics tool indicating the presence or absence of the biomarkers in the test sample. As an example application of the system, we will focus our effort on detection of a panel of 3 serum antibodies for human papillomavirus (HPV) as a means for identifying patients with greater susceptibility to oropharyngeal carcinoma (OPC), although the same technology can be extended to detection of broad classes of antibody biomarkers. The new instrument will be first tested upon biomarkers spiked into whole blood, with results compared against single-antibody ELISA in microplates and the Luminex bead-based system. Further validation and comparison will be performed upon clinical blood samples from patients with known HPV exposure. Our long-term goal is to demonstrate a prototype system that can be applied broadly for multiplexed serum antibody biomarker analysis.

Public Health Relevance

of the application The project aims to develop a highly sensitive, multiplexed platform technology for detection of serum antibodies that have been identified as biomarkers for viral infection, cancer, and autoimmune disease. Microfluidics and on-chip blood filtration are used to automate the assay process to obtain results in <60 minutes using a single droplet of blood.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM108584-03
Application #
9102244
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Edmonds, Charles G
Project Start
2014-06-01
Project End
2018-05-31
Budget Start
2016-06-01
Budget End
2017-05-31
Support Year
3
Fiscal Year
2016
Total Cost
$374,785
Indirect Cost
$51,341
Name
University of Illinois Urbana-Champaign
Department
Type
Organized Research Units
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Liang, Li-Guo; Kong, Meng-Qi; Zhou, Sherry et al. (2017) An integrated double-filtration microfluidic device for isolation, enrichment and quantification of urinary extracellular vesicles for detection of bladder cancer. Sci Rep 7:46224
Liang, Li-Guo; Sheng, Ye-Feng; Zhou, Sherry et al. (2017) An Integrated Double-Filtration Microfluidic Device for Detection of Extracellular Vesicles from Urine for Bladder Cancer Diagnosis. Methods Mol Biol 1660:355-364
Inan, Hakan; Wang, Shuqi; Inci, Fatih et al. (2017) Isolation, Detection, and Quantification of Cancer Biomarkers in HPV-Associated Malignancies. Sci Rep 7:3322
Sher, Mazhar; Zhuang, Rachel; Demirci, Utkan et al. (2017) Paper-based analytical devices for clinical diagnosis: recent advances in the fabrication techniques and sensing mechanisms. Expert Rev Mol Diagn 17:351-366
Inan, Hakan; Poyraz, Muhammet; Inci, Fatih et al. (2017) Photonic crystals: emerging biosensors and their promise for point-of-care applications. Chem Soc Rev 46:366-388
Ermis, Menekse; Akkaynak, Derya; Chen, Pu et al. (2016) A high throughput approach for analysis of cell nuclear deformability at single cell level. Sci Rep 6:36917
Wang, ShuQi; Lifson, Mark A; Inci, Fatih et al. (2016) Advances in addressing technical challenges of point-of-care diagnostics in resource-limited settings. Expert Rev Mol Diagn 16:449-59
El Assal, Rami; Gurkan, Umut A; Chen, Pu et al. (2016) 3-D Microwell Array System for Culturing Virus Infected Tumor Cells. Sci Rep 6:39144
Wang, ShuQi; Chinnasamy, Thiruppathiraja; Lifson, Mark A et al. (2016) Flexible Substrate-Based Devices for Point-of-Care Diagnostics. Trends Biotechnol 34:909-921
Tan, Yafang; Tang, Tiantian; Xu, Haisheng et al. (2015) High sensitivity automated multiplexed immunoassays using photonic crystal enhanced fluorescence microfluidic system. Biosens Bioelectron 73:32-40