Measurements of proteins are of central importance in biomedical research, and disease-related applications include early detection, causal identification, and treatment monitoring. Sensitive, specific, fast, inexpensive detection of multiple proteins in patient samples is critically important for future public health. The broad, long term goals of this project are to develop ultrasensitive new array devices for accurate measurements of panels of proteins. Recent research has shown that panels of 4 to 10 biomarker proteins, as opposed to the single biomarkers now used, will provide much more reliable cancer detection and monitoring. We will develop and optimize microfluidic arrays for proteins that are accurate, cheap, and can detect multiple proteins at levels 10-100 fold lower than any commercial alternatives. This project targets arrays for small panels of biomarker proteins in serum and tissue, not proteomic biomarker discovery or analysis. In the past funding period, we developed nanoparticle-based strategies for ultrasensitive protein immunoarrays using many thousands of enzyme labels on magnetic particles for electrochemical detection, or dye-nanoparticle labels for electro-optical detection. We utilized prototype sensors and arrays to detect up to four cancer biomarker proteins in the serum of prostate cancer patients with high accuracy. We recently combined nanostructured sensor arrays with microfluidics and off-line protein capture by the multilabel magnetic-antibody particles. This powerful approach greatly decreases interferences and can detect biomarker proteins in serum at levels of 100 fg mL-1 (~3 fM), 10-100 fold below capabilities of commercial assays. Such high sensitivity provides new opportunities for detection of biomarkers with inherently ultralow levels. This renewal project seeks to translate our ultrasensitive protein detection approaches into the realization of widespread clinical, surgical, and research applications. The deliverables are optimized, validated microfluidic array devices for ultrasensitive detection of virtually any small panel of proteins. A high-speed array for detection of cancer cell metastasis (spreading) to lymph nodes during cancer surgery will also be developed. For proof-of-concept, specific devices will be designed to predict the probability of oral cancer from collections of serum samples, and to clearly distinguish between viral (humanpapilloma virus) and non-viral oral cancers. Patient and control samples obtained by our NIDCR/NIH collaborators will be used to establish clinical sensitivity and selectivity of the immunoassay devices. Summarized specific aims are: (1) Optimize microfluidic systems featuring nanostructured sensor chips and multilabel amperometry for an 8-protein oral cancer panel;(2) Design and optimize a simple electro-optical microfluidic array using dye-nanoparticle labels detected by electrochemiluminescence (ECL);(3) Establish clinical sensitivity and selectivity for oral cancer diagnosis for the best array systems;(4) Develop a rapid microfluidic assay for detection of a metastasis biomarker in lymph nodes. 1

Public Health Relevance

Relevance to Public Health This project will develop ultrasensitive devices to detect collections of protein biomarkers in patient serum. It will provide new tools for reliable early diagnosis and monitoring of cancer to facilitate personalized therapy. It will also develop a rapid device to detect the spread (metastasis) of cancer.

National Institute of Health (NIH)
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Research Project (R01)
Project #
Application #
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Korte, Brenda
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Connecticut
Schools of Arts and Sciences
United States
Zip Code
Tang, C K; Vaze, A; Rusling, J F (2014) Paper-based electrochemical immunoassay for rapid, inexpensive cancer biomarker protein detection. Anal Methods 6:8878-8881
Joshi, Amit A; Peczuh, Mark W; Kumar, Challa V et al. (2014) Ultrasensitive carbohydrate-peptide SPR imaging microarray for diagnosing IgE mediated peanut allergy. Analyst 139:5728-33
Sharifi, Roholah; Samaraweera, Milinda; Gascón, José A et al. (2014) Thermodynamics of the quasi-epitaxial flavin assembly around various-chirality carbon nanotubes. J Am Chem Soc 136:7452-63
Rusling, James F; Bishop, Gregory W; Doan, Nhi et al. (2014) Nanomaterials and biomaterials in electrochemical arrays for protein detection. J Mater Chem B Mater Biol Med 2:
Otieno, Brunah A; Krause, Colleen E; Latus, Alina et al. (2014) On-line protein capture on magnetic beads for ultrasensitive microfluidic immunoassays of cancer biomarkers. Biosens Bioelectron 53:268-74
Guz, Nataliia; Halámek, Jan; Rusling, James F et al. (2014) A biocatalytic cascade with several output signals--towards biosensors with different levels of confidence. Anal Bioanal Chem 406:3365-70
Wang, Yixian; Kececi, Kaan; Mirkin, Michael V et al. (2013) Resistive-pulse measurements with nanopipettes: detection of Au nanoparticles and nanoparticle-bound anti-peanut IgY. Chem Sci 4:655-663
Mani, Vigneshwaran; Kadimisetty, Karteek; Malla, Spundana et al. (2013) Paper-based electrochemiluminescent screening for genotoxic activity in the environment. Environ Sci Technol 47:1937-44
Croce Jr, Robert A; Vaddiraju, SanthiSagar; Kondo, Jun et al. (2013) A miniaturized transcutaneous system for continuous glucose monitoring. Biomed Microdevices 15:151-60
Sardesai, Naimish P; Kadimisetty, Karteek; Faria, Ronaldo et al. (2013) A microfluidic electrochemiluminescent device for detecting cancer biomarker proteins. Anal Bioanal Chem 405:3831-8

Showing the most recent 10 out of 16 publications