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


Sadik D. A. , Eksi-Kocak H., ERTAŞ G., BOYACI İ. H. , MUTLU M.

SURFACE AND INTERFACE ANALYSIS, cilt.50, ss.866-878, 2018 (SCI İndekslerine Giren Dergi) identifier identifier

  • Cilt numarası: 50 Konu: 9
  • Basım Tarihi: 2018
  • Doi Numarası: 10.1002/sia.6489
  • Dergi Adı: SURFACE AND INTERFACE ANALYSIS
  • Sayfa Sayıları: ss.866-878

Özet

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.