Mixed-monolayer of N-hydroxysuccinimide-terminated cross-linker and short alkanethiol to improve the efficiency of biomolecule binding for biosensing

Date

2018-09-01

Author

Sadik, Demet Ataman
Eksi-Kocak, Haslet
Ertaş, Gülay
BOYACI, İSMAİL HAKKI
MUTLU, MEHMET

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The goal of this study was to use a novel surface chemistry for modifying gold surfaces to decrease the steric hindrance, minimize the nonspecific bindings while providing directed immobilization of proteins for advancing the transducer property and to provide a biosensing platform for surface plasmon resonance (SPR) applications. Mixed self-assembled monolayers (mSAMs) were prepared using 3,3-Dithiodipropionic acid di (N-hydroxysuccinimide ester) (DSP) and 6-mercapto-1-hexanol (MCH) and the selected model proteins bovine serum albumin (BSA) and lysozyme were tested for binding efficiency. First, binding of these two proteins at constant concentration to different DSP:MCH mSAMs were compared to deduce the best molar ratio for forming mSAM using a continuous flow system coupled to SPR. Coincidently the maximum protein binding DSP:MCH mSAM were the same for both proteins. The change in Response Unit (RU) signal due to protein binding between DSP SAM and maximum protein binding DSP:MCH mSAM for lysozyme binding was more in comparison to BSA binding. Second, the effect of BSA and lysozyme concentration on binding efficiency to maximum protein binding DSP:MCH mSAM were compared and discussed. Lysozyme and BSA were shown to reach saturations on the same monolayer at concentrations of 5.7x10(-5) and 8.96x10(-6) [M] respectively, hence the molar ratio for limit concentrations is 6:1. The DSP SAM, MCH SAM, and DSP:MCH mSAMs where maximum and minimum protein binding occurs were also characterized with XPS and Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy. Blank gold surface, maximum protein binding DSP:MCH mSAM and BSA immobilized DSP:MCH mSAM on gold surface were also investigated utilizing tapping mode AFM.

Subject Keywords

Gold surface modification, Transducer, Surface plasmon resonance (SPR), Surface characterization, Protein immobilization, Mixed self-assembled monolayer (mSAMs)

URI

https://hdl.handle.net/11511/37118

Journal

SURFACE AND INTERFACE ANALYSIS

DOI

https://doi.org/10.1002/sia.6489

Collections

Department of Chemistry, Article

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Citation Formats

D. A. Sadik, H. Eksi-Kocak, G. Ertaş, İ. H. BOYACI, and M. MUTLU, “Mixed-monolayer of N-hydroxysuccinimide-terminated cross-linker and short alkanethiol to improve the efficiency of biomolecule binding for biosensing,” SURFACE AND INTERFACE ANALYSIS, pp. 866–878, 2018, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/37118.