An immunoflorescence method for monitoring
a wastewater bioreactor
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Principal
Investigator
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James
Wang Norman
Chiu |
|
RET
Teachers
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Winnifred "Winnie" Eke |
| Research Abstract | Nitrogen reduction is becoming a critical task for many municipal wastewater treatment plants such as the Massachusetts Water Resources Authority’s (MWRA) Deer Island Treatment Plant. Biological nitrogen removal is a multi-step process carried out by bacteria of several trophic groups. The conventional reactor control is based on the chemical data collected from bulk liquid. Assumptions and estimate, such as diffusion coefficients and active biomass, have to be made in the process design and reactor control. Such approach represents an intrinsic limitation, especially when the treatment process includes multiple biological reactions involving diverse microbial populations. A molecular biology-based method targeting directly at selected microbial activities can lead to significantly improved design and operational control. The specific objective of this research is to develop an immunological method targeting a specific enzyme controlling the nitrogen removal process. |
| Research Experience | For immunoassay, microtiter-based competitive enzyme-linked immunoabsorbent assay (ELISA) will be used. In brief, fixed amount of a specific monoclonal antibody is first immobilized on the surface of 96-microtiter wells. A mixture that contains the sample of protein extract plus known amount of fluorescently-labeled nitrite oxidoreductase standard is added into microtiter wells. The nitrite oxidoreductase in the sample will compete with the labeled enzyme standard for limited binding sites on the immobilized antibodies. After the immunoreaction is completed, any unbound molecules are removed from the well. The amount of captured enzyme standard will be determined by measuring the signal from its fluorescent tag. By comparing to the fluorescent signal that is obtained from a control experiment that consists of zero nitrite oxidoreductase from the protein extract, the amount of nitrite oxidoreductase in the sample can be calculated. The results from this experiment will provide the information on the total amount of nitrite oxidoreductase in the protein extract. |
| Expectations of RET | The participant is expected to work jointly with graduate students in the laboratory. He/she will need to ultimately develop an understanding in microbial metabolism and the associated biochemical mechanisms. The participant is expected to develop the skills of preparing environmental samples for molecular biology analyses - cell extraction and purification, enzyme labeling, fluorescence measurement, and etc. The participant should ultimately be able to link the cell biology information to engineering treatment processes. |
| Helpful Skills or Interests | The participant should have an interest in basic biochemistry and microbiology and how they are applied in environmental engineering processes. |
