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

被引:17
作者
Chamas, Alexandre [1 ]
Ha Thi Minh Pham [1 ,2 ]
Jaehne, Martin [3 ]
Hettwer, Karina [3 ]
Uhlig, Steffen [3 ]
Simon, Kirsten [4 ]
Einspanier, Almuth [5 ]
Baronian, Kim [6 ]
Kunze, Gotthard [1 ]
机构
[1] Leibniz Inst Plant Genet & Crop Plant Res IPK, Corrensstr 3, D-06466 Gatersleben, Germany
[2] Vietnam Acad Sci & Technol, Inst Biotechnol, Cau Giay, Ha Noi, Vietnam
[3] QuoData GmbH, Dresden, Germany
[4] New Diagnost GmbH, Freising Weihenstephan, Germany
[5] Inst Physiol Chem, Fac Vet Med, Leipzig, Germany
[6] Univ Canterbury, Sch Biol Sci, Christchurch, New Zealand
关键词
hormone assay; Arxula adeninivorans; fluorescence; marmoset serum; ARXULA-ADENINIVORANS CELLS; ENDOCRINE DISRUPTORS; WASTE-WATER; ESTROGENIC COMPOUNDS; FLUORESCENT PROTEIN; CALLITHRIX-JACCHUS; MARMOSET MONKEY; IN-VITRO; ASSAY; PROGESTERONE;
D O I
10.1002/bit.26249
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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. (c) 2017 Wiley Periodicals, Inc.
引用
收藏
页码:1539 / 1549
页数:11
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