Biosensing estrogenic endocrine disruptors in human blood and urine: A RAPID cell-free protein synthesis approach.

Autor: Salehi ASM; Department of Chemical Engineering, Brigham Young University, Provo, UT, USA., Yang SO; Department of Chemical Engineering, Brigham Young University, Provo, UT, USA., Earl CC; Department of Chemical Engineering, Brigham Young University, Provo, UT, USA., Shakalli Tang MJ; Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, USA., Porter Hunt J; Department of Chemical Engineering, Brigham Young University, Provo, UT, USA., Smith MT; Department of Chemical Engineering, Brigham Young University, Provo, UT, USA., Wood DW; Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, USA. Electronic address: wood.750@osu.edu., Bundy BC; Department of Chemical Engineering, Brigham Young University, Provo, UT, USA. Electronic address: bundy@byu.edu.
Jazyk: angličtina
Zdroj: Toxicology and applied pharmacology [Toxicol Appl Pharmacol] 2018 Apr 15; Vol. 345, pp. 19-25. Date of Electronic Publication: 2018 Feb 27.
DOI: 10.1016/j.taap.2018.02.016
Abstrakt: Many diseases and disorders are linked to exposure to endocrine disrupting chemicals (EDCs) that mimic the function of natural estrogen hormones. Here we present a Rapid Adaptable Portable In-vitro Detection biosensor platform (RAPID) for detecting chemicals that interact with the human estrogen receptor β (hERβ). This biosensor consists of an allosteric fusion protein, which is expressed using cell-free protein synthesis technology and is directly assayed by a colorimetric response. The resultant biosensor successfully detected known EDCs of hERβ (BPA, E2, and DPN) at similar or better detection range than an analogous cell-based biosensor, but in a fraction of time. We also engineered cell-free protein synthesis reactions with RNAse inhibitors to increase production yields in the presence of human blood and urine. The RAPID biosensor successfully detects EDCs in these human samples in the presence of RNAse inhibitors. Engineered cell-free protein synthesis facilitates the use of protein biosensors in complex sample matrices without cumbersome protein purification.
(Copyright © 2018. Published by Elsevier Inc.)
Databáze: MEDLINE
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