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

Kód výsledku v IS VaVaI

RIV/00216224:14310/19:00107427 - isvavai.cz

Výsledek na webu

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 - Digital Object Identifier

10.1177/0003702819826280

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

21100 - Other engineering and technologies

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

APPLIED SPECTROSCOPY

ISSN

0003-7028

e-ISSN

1943-3530

Svazek periodika

73

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

10

Strana od-do

261-270

Kód UT WoS článku

000460535500002

EID výsledku v databázi Scopus

2-s2.0-85061963600