The binding potency of those dust extracts was only slightly lower than the positive control (12.5 μM of rosiglitazone), which could completely inhibit the binding between the PPARγ LBD and PPARγ Green probe.
It might be possible that compounds in dust, such as TBB, TBPH, PBDEs, and DEHP, could be metabolized to PPARγ-active ligands after incubation, which was supported by the increased binding potency of the prepared MIX containing these chemicals.
To test this possibility we compared VNTR domain sizes with DC-SIGN binding potency of the corresponding milks.
To quantitatively observe the change with different doses, we conducted a dose response analysis to investigate the binding potency of the MIX, bioactivated MIX, and SRM 2585.
Overall, we conclude that the FP assay was appropriate and efficient to evaluate the binding potency of the tested compounds and dust extracts.
In addition, the binding potency of the house dust in our study might be underestimated due to FP interference from the dust matrix.
19 The increase in binding potency of the phenylpropyl group can also been seen in the increase in potency when added to the clomipramine (E) or chlorpromazine (H) scaffolds.
Briefly, we used a commercially available high-throughput ligand binding assay (PolarScreen™ PPARγ-Competitor Assay Kit; Invitrogen, Carlsbad, CA) to investigate the binding potency of the tested compounds to PPARγ LBD.
Following GPC cleanup and dilution, a single concentration of 3 mg dust equivalent quantity (DEQ) per milliliter PPARγ assay medium was prepared to qualitatively investigate the relative PPARγ binding potency of the dust samples; we examined the full dose response of one potent dust extract.
Here, it is demonstrated that more data on TTR-binding compounds is needed to cover the potential binding potency of compounds without these typical characteristics.
