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Bioluminescence Inhibition Assay for the Detection of Hydroxylated Polychlorinated Biphenyls
journal contribution
posted on 2012-09-18, 00:00 authored by Krystal Teasley Hamorsky, C. Mark Ensor, Emre Dikici, Patrizia Pasini, Leonidas Bachas, Sylvia DaunertHydroxylated polychlorinated biphenyls (OH-PCBs) are
an important class of contaminants that mainly originate from polychlorinated
biphenyl metabolism. They may conceivably be as dangerous and persistent
as the parent compounds; most prominently, OH-PCBs are endocrine disruptors.
Due to increasing evidence of the presence of OH-PCBs in the environment
and in living organisms, including humans, and of their toxicity,
methods of detection for OH-PCBs are needed in the environmental and
medical fields. Herein, we describe the development and optimization
of a protein-based inhibition assay for the quantification of OH-PCBs.
Specifically, the photoprotein aequorin was utilized for the detection
of OH-PCBs. We hypothesized that OH-PCBs interact with aequorin, and
we established that OH-PCBs actually inhibit the bioluminescence of
aequorin in a dose-dependent manner. We took advantage of this phenomenon
to develop an assay that is capable of detecting a wide variety of
OH-PCBs with a range of detection limits, the best detection limit
being 11 nM for the compound 2-hydroxy-2′,3,4′,5′,6-pentachorobiphenyl.
The viability of this system for the screening of OH-PCBs in spiked
biological and environmental samples was also established. We envision
the implementation of this novel bioluminescence inhibition assay
as a rapid, sensitive, and cost-effective method for monitoring OH-PCBs.
Furthermore, to the best of our knowledge, this is the first time
aequorin has been employed to detect an analyte by the inhibition
of its bioluminescence reaction. Hence, this strategy may prove to
be a general approach for the development of a new generation of protein-based
inhibition assays.