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We described the rapid production of the domain III (DIII) of the envelope (E) protein in plants as a vaccine candidate for West Nile Virus (WNV). Using various combinations of vector modules of a deconstructed viral vector expression system, DIII was produced in three subcellular compartments in leaves of Nicotiana

We described the rapid production of the domain III (DIII) of the envelope (E) protein in plants as a vaccine candidate for West Nile Virus (WNV). Using various combinations of vector modules of a deconstructed viral vector expression system, DIII was produced in three subcellular compartments in leaves of Nicotiana benthamiana by transient expression. DIII expressed at much higher levels when targeted to the endoplasmic reticulum (ER) than that targeted to the chloroplast or the cytosol, with accumulation level up to 73 μg DIII per gram of leaf fresh weight within 4 days after infiltration. Plant ER-derived DIII was soluble and readily purified to > 95% homogeneity without the time-consuming process of denaturing and refolding. Further analysis revealed that plant-produced DIII was processed properly and demonstrated specific binding to an anti-DIII monoclonal antibody that recognizes a conformational epitope. Furthermore, subcutaneous immunization of mice with 5 and 25 μg of purified DIII elicited a potent systemic response. This study provided the proof of principle for rapidly producing immunogenic vaccine candidates against WNV in plants with low cost and scalability.

ContributorsHe, Junyun (Author) / Peng, Li (Author) / Lai, Huafang (Author) / Hurtado, Jonathan (Author) / Stahnke, Jake (Author) / Chen, Qiang (Author) / ASU Biodesign Center Immunotherapy, Vaccines and Virotherapy (Contributor) / Biodesign Institute (Contributor)
Created2014-04-03
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Description

There is an increasing awareness that health care must move from post-symptomatic treatment to presymptomatic intervention. An ideal system would allow regular inexpensive monitoring of health status using circulating antibodies to report on health fluctuations. Recently, we demonstrated that peptide microarrays can do this through antibody signatures (immunosignatures). Unfortunately, printed

There is an increasing awareness that health care must move from post-symptomatic treatment to presymptomatic intervention. An ideal system would allow regular inexpensive monitoring of health status using circulating antibodies to report on health fluctuations. Recently, we demonstrated that peptide microarrays can do this through antibody signatures (immunosignatures). Unfortunately, printed microarrays are not scalable. Here we demonstrate a platform based on fabricating microarrays (~10 M peptides per slide, 330,000 peptides per assay) on silicon wafers using equipment common to semiconductor manufacturing. The potential of these microarrays for comprehensive health monitoring is verified through the simultaneous detection and classification of six different infectious diseases and six different cancers. Besides diagnostics, these high-density peptide chips have numerous other applications both in health care and elsewhere.

ContributorsLegutki, Joseph Barten (Author) / Zhao, Zhan-Gong (Author) / Greving, Matt (Author) / Woodbury, Neal (Author) / Johnston, Stephen (Author) / Stafford, Phillip (Author) / Biodesign Institute (Contributor)
Created2014-09-03
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Description

We present a new method of chemical quantification utilizing thermal analysis for the detection of relative humidity. By measuring the temperature change of a hydrophilically-modified temperature sensing element vs. a hydrophobically-modified reference element, the total heat from chemical interactions in the sensing element can be measured and used to calculate

We present a new method of chemical quantification utilizing thermal analysis for the detection of relative humidity. By measuring the temperature change of a hydrophilically-modified temperature sensing element vs. a hydrophobically-modified reference element, the total heat from chemical interactions in the sensing element can be measured and used to calculate a change in relative humidity. We have probed the concept by assuming constant temperature streams, and having constant reference humidity (~0% in this case). The concept has been probed with the two methods presented here: (1) a thermistor-based method and (2) a thermographic method. For the first method, a hydrophilically-modified thermistor was used, and a detection range of 0–75% relative humidity was demonstrated. For the second method, a hydrophilically-modified disposable surface (sensing element) and thermal camera were used, and thermal signatures for different relative humidity were demonstrated. These new methods offer opportunities in either chemically harsh environments or in rapidly changing environments. For sensing humidity in a chemically harsh environment, a hydrophilically-modified thermistor can provide a sensing method, eliminating the exposure of metallic contacts, which can be easily corroded by the environment. On the other hand, the thermographic method can be applied with a disposable non-contact sensing element, which is a low-cost upkeep option in environments where damage or fouling is inevitable. In addition, for environments that are rapidly changing, the thermographic method could potentially provide a very rapid humidity measurement as the chemical interactions are rapid and their changes are easily quantified.

ContributorsBridgeman, Devon (Author) / Tsow, Francis (Author) / Xian, Xiaojun (Author) / Chang, Qinan (Author) / Liu, Yongming (Author) / Forzani, Erica (Author) / Biodesign Institute (Contributor)
Created2017-05-24