Fabrication of dense, uniform aminosilane monolayers: A platform for protein or ligand immobilization

Sanjukta Guha Thakurta, Anuradha Subramanian

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Designing biointerfaces with enhanced biomolecule binding abilities, along with their distribution and presentation on the surface, largely requires control over underlying surface chemistries. In an effort to correlate experimental parameters to surface properties, ligand distribution and biomolecule binding, the influence of (3-aminopropyl)triethoxysilane (APTES) concentration (1, 2 and 4vol%) and duration of the silanization reaction (5-60min) on the number of available NH2 groups were determined and correlated with the amount of surface bound human immunoglobulin G (HIgG). Surfaces silanized with 2vol% APTES for 30min yielded a densely populated silane monolayer (1.0-1.2nm) where the average molecular orientation was 38±2° with respect to the surface normal. A surface density of NH2 moieties of ~1015/cm2 was obtained and was significantly higher compared to other conditions evaluated. The combined data from ellipsometry and atomic force microscopy (AFM) analyses supports our findings. Coupling of HIgG to surfaces silanized with 2vol% APTES for 30min yielded ~1013IgG/cm2, which was significantly higher than values obtained at 1vol% APTES for 30min. The distribution of immobilized HIgG was noted to be dense and uniform on surfaces silanized with 2vol% APTES when visualized using Nanogold® antibody conjugates. This study highlights the critical role of the experimental parameters that impact the biomolecule immobilization process on functionalized surfaces.

Original languageEnglish (US)
Pages (from-to)384-392
Number of pages9
JournalColloids and Surfaces A: Physicochemical and Engineering Aspects
Volume414
DOIs
StatePublished - Nov 20 2012

Keywords

  • 3-Aminopropyltriethoxysilane
  • Antibody conjugation
  • Biosensor
  • Self-assembled monolayer
  • Silanization
  • Surface characterization
  • Surface topography

ASJC Scopus subject areas

  • Surfaces and Interfaces
  • Physical and Theoretical Chemistry
  • Colloid and Surface Chemistry

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