Publication details

Treatment of Surface Plasmon Resonance (SPR) Background in Total Internal Reflection Ellipsometry: Characterization of RNA Polymerase II Film Formation

Authors

HEMZAL Dušan KANG Yu Ri DVOŘÁK Jan KABZINSKI Tomasz KUBÍČEK Karel YOUNG DONG Kim HUMLÍČEK Josef

Year of publication 2019
Type Article in Periodical
Magazine / Source APPLIED SPECTROSCOPY
MU Faculty or unit

Faculty of Science

Citation
Web https://journals.sagepub.com/doi/abs/10.1177/0003702819826280?rfr_dat=cr_pub%3Dpubmed&url_ver=Z39.88-2003&rfr_id=ori%3Arid%3Acrossref.org&journalCode=aspc
Doi http://dx.doi.org/10.1177/0003702819826280
Keywords Spectroscopic ellipsometry; surface plasmon resonance; SPR; total internal reflection ellipsometry; TIRE; RNA polymerase II; C-terminal domain; CTD; Rtt103 protein
Description To deal with the general problem of biomolecule specific binding analysis, we have applied the technique of difference spectra to the surface plasmon resonance (SPR)-enhanced total internal reflection ellipsometry measurement. We suggest a three-step treatment of the SPR background that can easily be integrated with the usual measurement routine. First, making use of the difference spectrum in ellipsometric angle Delta, single peak footprints of the topmost layer are obtained that facilitate its sensitive detection during film growth. Subsequently, circumventing the need for explicit knowledge of the substrate properties, the difference spectra peaks can be used for the end-point analysis of a binding. Finally, tracking the binding effectivity of the analyte we determine the injection speed and analyte concentration windows needed for successful monitoring of the film growth. We demonstrate our approach on a comprehensive two-stage binding experiment involving two biologically relevant molecules: the C-terminal domain (CTD) of RNA polymerase II and CTD-interacting domain of one of its transcription factors, the Rtt103 protein.
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