Simultaneous detection of three sex steroid hormone classes using a novel yeast‐based biosensor

Abstract

A biosensor detecting estrogens, progestogens, and androgens in complex samples and in a single step is described. Three Arxula adeninivorans yeast strains were created, each strain producing a different recombinant human hormone receptor and a different fluorescent reporter protein. These strains were then mixed to create G1212/YRC102-hHR-fluo, the biological component of the biosensor. During incubation with G1212/YRC102-hHR-fluo, hormones present in a sample bind to their target receptor, which leads to the production of a specific fluorescent protein. Three fluorescence scans of the yeast suspension determine which fluorescence protein has been produced, thus revealing which hormone receptor (estrogen, progesterone, and androgen) has been activated by the hormones or hormone mimics present in the sample. The biosensor has similar sensitivities to the existing A. adeninivorans cell-based assays. The detection of the three hormone classes in one single experiment reduces the labor and time required to assay for the three hormone classes. The biosensor was also trialed with animal serum samples for the detection of progestogens, androgens, and estrogens and gave results that correlated well with ELISA analysis in case of progestogens. These results highlight the potential usefulness of the biosensor for comprehensive determination of hormone status in samples from veterinary origin. Biotechnol. Bioeng. 2017;114: 1539-1549. © 2017 Wiley Periodicals, Inc.

Estrogenic activity of biological samples as a biomarker

“…Other authors applied extraction to biological samples before the analysis: in the studies of Pedersen et al (2010) and Chamas et al (2017) samples were extracted using a liquid-liquid extraction with methyl tert-butyl ether and diethyl ether respectively, while in the study of Kanaya et al (2015) samples were extracted using a solid-phase extraction.…”

Recent trends in biocatalysis

“… 151 More recently, Chamas et al created biosensors for the detection of estrogens, progestogens (see also Scheme 8 ), and androgens in Arxula adeninivorans yeast strains by coupling human hormone receptors and different fluorescent reporter proteins. 152 A complementary approach was followed by Maser and Xiong and put into perspective of alternative steroid-sensing methods. 153 Their Comamonas testosteroni steroid-sensor (COSS) system is based on the insertion of a green fluorescent protein ( gfp ) gene upstream of the regulatory region of the hsdA gene encoding a 3α-hydroxysteroid dehydrogenase/carbonyl reductase.…”

A fluorescence‐based yeast sensor for monitoring acetic acid

“…Very recently a microfluidic flow-through device containing genetically modified yeast cells for the detection of diclofenac in wastewater has been reported [14]. Yeast cells have been used to sense a large variety of analytes including glucose [15,16], copper ions [17][18][19], hormonally active substances [20][21][22], toxic substances [23][24][25] as well asantibiotics [26] or analgesics [27].…”

“…20 min after activation of the pheromone signal cascade. Consequently, the R-cells generate a strong fluorescence signal and co-cultivation of S-and R-cells is an appropriate way A B F I G U R E 5 (A) Scheme for using a yeast pheromone-based system for signal amplification.…”