Bioconjugation of the estrogen receptor hERα to a quantum dot dye for a controlled immobilization on a SiO2 surface

We investigated the immobilization of the estrogen receptor hERα on silanized SiO2 surfaces for biosensor applications. The conjugation of the estrogen receptor hERα to the quantum dot dye QD655 was achieved. In order to obtain an optimal immobilization of the estrogen receptor hERα on the functionalized SiO2 surface, the bioconjugate hERα-QD655 (Rcpt-qd655) solution was prepared with a higher molar ratio of 10–15 between the QDs and the receptors. A blue laser with an excitation wavelength of 405 nm was used for photoluminescence spectroscopy (PL) investigations to monitor the bioconjugate Rcpt-qd655 immobilization on the silanized SiO2 surfaces with three different functional groups, namely NH2, -COO-, and –SH. Several wash processes were applied to remove the excess receptors from the surface after the immobilization. A Fourier transform infrared spectroscopy (FTIR) was used to control the biofilm background after each wash of the receptor-coated surface which allows the optimization of the immobilization protocol. In order to test its stability a quartz crystal microbalance (QCM) was employed and the receptor density was calculated. Finally the optimal biolayer (silane film + hERα receptor) was tested for measurements of 17ß-estradiol (E2) with a concentration of 1 μM in waterish solution. The measurement concept outlined in [L. Rebohle et al., Vacuum 83 (2009) 24–28] was applied. The whole system was investigated by PL, which exhibits two color signals, namely from the receptor and the detected E2 molecules.

This paper demonstrated the immobilization of the conjugated estrogen receptor hERα with QDS655 dye on the silanized SiO2 surfaces. The immobilization of the estrogen receptor on the silanized SiO2 surface occurred under a certain angle which makes the binding pocket of the receptor accessible for estrogen docking.Figure optionsDownload high-quality image (72 K)Download as PowerPoint slideResearch highlights
► The conjugation of the estrogen receptor hERα to the quantum dots dye QD655 and their immobilization on the three different silanized SiO2 surfaces.
► A higher molarity ratio of 10–20 between the QDs and the receptors leads to the immobilization of the estrogen receptor on the silanized SiO2 surface under a certain angle.
► The binding pocket of the receptor is accessible for estrogen docking.
► Detection of 17ß-estradiol (E2) concentration in waterish solution by use of a second color of dye QD800 at 800 nm.