| Title: | Hydrophobin-Based Surface Engineering for Sensitive and Robust Quantification of Yeast Pheromones |
| Author(s): | Hennig S; Rodel G; Ostermann K; |
| Address: | "Institute of Genetics, Technische Universitat Dresden, 01062 Dresden, Germany. Stefan.Hennig1@tu-dresden.de. Institute of Genetics, Technische Universitat Dresden, 01062 Dresden, Germany. Gerhard.Roedel@tu-dresden.de. Institute of Genetics, Technische Universitat Dresden, 01062 Dresden, Germany. Kai.Ostermann@tu-dresden.de" |
| ISSN/ISBN: | 1424-8220 (Electronic) 1424-8220 (Linking) |
| Abstract: | "Detection and quantification of small peptides, such as yeast pheromones, are often challenging. We developed a highly sensitive and robust affinity-assay for the quantification of the alpha-factor pheromone of Saccharomyces cerevisiae based on recombinant hydrophobins. These small, amphipathic proteins self-assemble into highly stable monolayers at hydrophilic-hydrophobic interfaces. Upon functionalization of solid supports with a combination of hydrophobins either lacking or exposing the alpha-factor, pheromone-specific antibodies were bound to the surface. Increasing concentrations of the pheromone competitively detached the antibodies, thus allowing for quantification of the pheromone. By adjusting the percentage of pheromone-exposing hydrophobins, the sensitivity of the assay could be precisely predefined. The assay proved to be highly robust against changes in sample matrix composition. Due to the high stability of hydrophobin layers, the functionalized surfaces could be repeatedly used without affecting the sensitivity. Furthermore, by using an inverse setup, the sensitivity was increased by three orders of magnitude, yielding a novel kind of biosensor for the yeast pheromone with the lowest limit of detection reported so far. This assay was applied to study the pheromone secretion of diverse yeast strains including a whole-cell biosensor strain of Schizosaccharomyces pombe modulating alpha-factor secretion in response to an environmental signal" |
| Keywords: | *Biosensing Techniques Hydrophobic and Hydrophilic Interactions Peptides *Pheromones Proteins *Saccharomyces cerevisiae analyte detection biosensor hydrophobin surface functionalization whole-cell biosensor yeast pheromone; |
| Notes: | "MedlineHennig, Stefan Rodel, Gerhard Ostermann, Kai eng Switzerland 2016/04/30 Sensors (Basel). 2016 Apr 27; 16(5):602. doi: 10.3390/s16050602" |
