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Quantitative determination of testosterone levels with biolayer interferometry.

Hao Zhang1, Wei Li1, Hong Luo2

  • 1School of Life Science and Technology, Changchun University of Science and Technology, 130022 Changchun, Jilin Province, PR China.

Chemico-Biological Interactions
|May 24, 2017
PubMed
Summary

A novel biosensor uses biolayer interferometry to detect environmental steroid hormones like testosterone. This method rapidly and accurately quantifies testosterone levels without labels, offering a new tool for environmental monitoring.

Keywords:
Biolayer interferometryComamonas testosteroniDNA/protein interactionQuantitative determination of testosterone

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Area of Science:

  • Environmental Science
  • Biotechnology
  • Analytical Chemistry

Background:

  • Steroid hormones are widespread environmental pollutants with endocrine-disrupting activities, posing risks to human health even at low concentrations.
  • Existing detection methods for environmental steroid hormones often require labels and can be time-consuming or complex.

Purpose of the Study:

  • To develop a novel, label-free biosensor system for the rapid and quantitative detection of steroid hormones in the environment.
  • To utilize biolayer interferometry for monitoring molecular interactions involved in steroid hormone detection.

Main Methods:

  • Development of a biosensor based on biolayer interferometry using optical fiber sensors.
  • Surface immobilization of double-stranded DNA fragments (OP1 and OP2) from Comamonas testosteroni.
  • Detection of repressor protein RepA binding to DNA and the influence of testosterone on this interaction.

Main Results:

  • The biosensor successfully detected DNA-protein interactions with high specificity and affinity, yielding dissociation constants for OP1 and OP2 with RepA.
  • RepA-testosterone binding was shown to alter RepA conformation, affecting its DNA binding affinity.
  • A quantitative correlation between detected signal and testosterone concentration was established (R² ≈ 0.97), enabling detection within a range of 2.13–136.63 ng/ml in 17 minutes.

Conclusions:

  • The developed biolayer interferometry biosensor system provides a rapid, quantitative, and label-free method for environmental testosterone detection.
  • The specific interaction between double-stranded OP1 DNA and RepA protein, modulated by testosterone, forms the basis for this sensitive detection technique.
  • This approach offers a promising tool for environmental monitoring of steroid hormone pollutants.