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Featured researches published by Bernard Munge.


Angewandte Chemie | 2011

Nanostructured Immunosensor for Attomolar Detection of Cancer Biomarker Interleukin-8 Using Massively Labeled Superparamagnetic Particles

Bernard Munge; Amy L. Coffey; Jaimee Doucette; Brian Somba; Vyomesh Patel; J. Silvio Gutkind; James F. Rusling

Extremely sensitive and accurate clinical measurements of biomarker proteins for early detection and monitoring of cancer pose a formidable challenge. However, successful inexpensive devices for reliable on-the-spot cancer diagnosis promise to lead to improved therapeutic outcomes with lower cost, decreased patient stress, and new targeted therapies.[1–3] Such devices will also provide tools for a better fundamental understanding of disease progression, and enable biomarker-based monitoring of therapy.[4] Herein, we report an ultra-sensitive immunosensor based on a glutathione-protected gold nanoparticle (GSH-AuNP) sensor surface. When combined with novel massively labeled paramagnetic particles for the electrochemical detection of cancer biomarker interleukin 8 (IL-8), we obtained an unprecedented detection limit (DL) of 1 fgmL−1 (100 aM) for IL-8, the lowest protein level yet detected in serum. Accuracy was demonstrated by determining IL-8 in conditioned media from head and neck squamous cell carcinoma (HNSCC) cells.


Analytical Chemistry | 2012

Ultrasensitive Detection of Cancer Biomarkers in the Clinic by Use of a Nanostructured Microfluidic Array

Vyomesh Patel; Bhaskara V. Chikkaveeraiah; Bernard Munge; Sok Ching Cheong; Rosnah Binti Zain; Mannil Thomas Abraham; Dipak K. Dey; J. Silvio Gutkind; James F. Rusling

Multiplexed biomarker protein detection holds unrealized promise for clinical cancer diagnostics due to lack of suitable measurement devices and lack of rigorously validated protein panels. Here we report an ultrasensitive electrochemical microfluidic array optimized to measure a four-protein panel of biomarker proteins, and we validate the protein panel for accurate oral cancer diagnostics. Unprecedented ultralow detection into the 5-50 fg·mL(-1) range was achieved for simultaneous measurement of proteins interleukin 6 (IL-6), IL-8, vascular endothelial growth factor (VEGF), and VEGF-C in diluted serum. The immunoarray achieves high sensitivity in 50 min assays by using off-line protein capture by magnetic beads carrying 400,000 enzyme labels and ~100,000 antibodies. After capture of the proteins and washing to inhibit nonspecific binding, the beads are magnetically separated and injected into the array for selective capture by antibodies on eight nanostructured sensors. Good correlations with enzyme-linked immunosorbent assays (ELISA) for protein determinations in conditioned cancer cell media confirmed the accuracy of this approach. Normalized means of the four protein levels in 78 oral cancer patient serum samples and 49 controls gave clinical sensitivity of 89% and specificity of 98% for oral cancer detection, demonstrating high diagnostic utility. The low-cost, easily fabricated immunoarray provides a rapid serum test for diagnosis and personalized therapy of oral cancer. The device is readily adaptable to clinical diagnostics of other cancers.


ChemBioChem | 2003

Optimization of Electrochemical and Peroxide-Driven Oxidation of Styrene with Ultrathin Polyion Films Containing Cytochrome P450cam and Myoglobin

Bernard Munge; Carmelita Estavillo; John B. Schenkman; James F. Rusling

The catalytic and electrochemical properties of myoglobin and cytochrome P450cam in films constructed with alternate polyion layers were optimized with respect to film thickness, polyion type, and pH. Electrochemical and hydrogen peroxide driven epoxidation of styrene catalyzed by the proteins was used as the test reaction. Ionic synthetic organic polymers such as poly(styrene sulfonate), as opposed to SiO2 nanoparticles or DNA, supported the best catalytic and electrochemical performance. Charge transport involving the iron heme proteins was achieved over 40–320 nm depending on the polyion material and is likely to involve electron hopping facilitated by extensive interlayer mixing. However, very thin films (ca. 12–25 nm) gave the largest turnover rates for the catalytic epoxidation of styrene, and thicker films were subject to reactant transport limitations. Classical bell‐shaped activity/pH profiles and turnover rates similar to those obtained in solution suggest that films grown layer‐by‐layer are applicable to turnover rate studies of enzymes for organic oxidations. Major advantages include enhanced enzyme stability and the tiny amount of protein required.


Faraday Discussions | 2000

Applications of polyion films containing biomolecules to sensing toxicity

James F. Rusling; Liping Zhou; Bernard Munge; Jing Yang; Carmelita Estavillo; John B. Schenkman

This paper describes several applications of polyion-biomolecule films on electrodes related to future development of in vitro chemical toxicity sensors. In the first example, composite films of DNA and ionomers cast onto pyrolytic graphite (PG) electrodes are shown to be useful for detecting DNA damage during incubation with the carcinogen styrene oxide at pH 5.5. Single electrodes can be used to estimate relative damage rates by derivative square wave voltammetry. Films containing the ionomer Nafion gave better reproducibility than another ionomer, Eastman AQ38. In the second example, films containing redox proteins myoglobin (Mb) and cytochrome (cyt) P450cam were constructed in alternate layers with polyions including DNA on rough PG electrodes. Films with reversible protein FeIII/FeII electrochemistry with up to 7 electroactive layers were made. Amounts of electroactive protein on rough PG that were 7 to 17-fold larger than in similar films on smooth gold were achieved because many more layers were electroactive. Films of Mb/DNA also showed oxidation peaks after short incubations with styrene oxide that may be attributable to DNA damage. Results are relevant to the future design of enzyme-DNA films which convert pollutants and drugs to reactive metabolites, followed by electrochemical detection of the resulting DNA damage.


Bioelectrochemistry | 2001

Electrochemical reactions of redox cofactors in Rhodobacter sphaeroides reaction center proteins in lipid films

Bernard Munge; Zeus Pendon; Harry A. Frank; James F. Rusling

Cyclic voltammetry of thin films made from the lipid dimyristoylphosphatidyl choline and reaction centers from the purple bacterium Rhodobacter sphaeroides on pyrolytic graphite electrodes in bromide-free pH 8 buffers at 4 degrees C revealed an oxidation peak at 0.98 V and a reduction peak at -0.17 V vs. NHE. No reverse CV peaks were found, suggesting chemical irreversibility. The reduction peak disappeared for reaction centers depleted of quinones, suggesting that the peak represents reduction of this cofactor. The oxidation peak showed a catalytic current increase in the presence of small amounts of ferrous cytochrome c, and decreased by 85% when illuminated by visible light, suggesting assignment to the primary donor (P) cofactor. While oxidized primary donor P(+) is destroyed upon electrochemical formation in the film, reaction of ferrous cyt c with P(+) suggests its persistence in the films on the microsecond time scale.


Archive | 2013

Nanoscience-Based Electrochemical Sensors and Arrays for Detection of Cancer Biomarker Proteins

James F. Rusling; Bernard Munge; Naimish P. Sardesai; Bhaskara V. Chikkaveeraiah

Measurement of panels of biomarker proteins in serum, tissue or saliva holds great promise for future cancer diagnostics. Broad implementation of this approach in the clinic requires new, low cost devices for multiplexed protein detection. Advanced nanomaterials coupled with electrochemical detection have provided new opportunities for development of such devices. This chapter reviews recent research in using nanoparticle labels and multiplexed detection in protein immunosensors. It focuses in part on research in our own laboratories on ultrasensitive protein immunosensors combining nanostructured electrodes with detection particles with up to 500,000 labels that detect as little as 1 fg/mL protein in diluted serum. Our most mature multiple protein detection arrays are multiplexed microfluidic devices with 8-nanostructured sensors utilizing massively labeled magnetic particles or polymers. This approach provides reliable detection for multiple proteins at levels well below 1 pg/mL, and shows excellent correlation with referee methods. The importance of validating panels of biomarkers for reliable cancer diagnostics is also stressed.


wearable and implantable body sensor networks | 2006

Single-walled carbon-nanotube forest immunosensor for amplified detection of cancer biomarkers

Joseph D. Gong; Gary C. Jensen; Ashwin Bhirde; Xin Yu; Bernard Munge; Voymesh Patel; SangNyon Kim; J. Silvio Gutkind; Fotios Papadimitrakopoulos; James F. Rusling

Prostate specific antigen (PSA) is a major biomarker used clinically in the detection of prostate cancer. Current commercial immunoassays for detection of this biomarker rely on spectroscopic absorbance and electro-chemiluminescence which are able to detect as low as 0.03 ng/mL of PSA within 100muL of serum. In this paper, a novel amplified electrochemical technique for detection of prostate specific antigen is described. This electrochemical approach is a highly sensitive method that can have detection limit of 0.004 ng/mL in only 10 mL of serum, a mass detection limit 75 times lower than current commercial techniques. This highly sensitive, low cost and fast detection method not only allows for detection of cancer biomarkers in serum, but also in tissue and cells. Moreover, immunoassay arrays based on this methodology will be highly promising for application in large-scale clinical screening and point-of-care diagnostics. Characterization of the anti-PSA amplification tag in this amplification system using TEM, AFM, SEM, and CE is also described


Journal of the American Chemical Society | 2006

Carbon Nanotube Amplification Strategies for Highly Sensitive Immunodetection of Cancer Biomarkers

Xin Yu; Bernard Munge; Vyomesh Patel; Gary C. Jensen; Ashwin Bhirde; Joseph D. Gong; Sang N. Kim; J. P. Gillespie; J. Silvio Gutkind; Fotios Papadimitrakopoulos; James F. Rusling


Analytical Chemistry | 2005

Multiple enzyme layers on carbon nanotubes for electrochemical detection down to 80 DNA copies.

Bernard Munge; Guodong Liu; and Greg E. Collins; Joseph Wang


Langmuir | 2000

Films of Manganese Oxide Nanoparticles with Polycations or Myoglobin from Alternate-Layer Adsorption†

Yuri Lvov; Bernard Munge; Oscar Giraldo; Izumi Ichinose; Steven L. Suib, ,§ and; James F. Rusling

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James F. Rusling

National Institutes of Health

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Vyomesh Patel

National Institutes of Health

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Xin Yu

University of Connecticut

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Ashwin Bhirde

University of Connecticut

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Gary C. Jensen

University of Connecticut

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Harry A. Frank

University of Connecticut

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Joseph D. Gong

University of Connecticut

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