Immobilization of oriented protein molecules on poly(ethylene glycol)-coated Si(111)

Taewoon Cha, Athena Guo, Yongseok Jun, Duanquing Pei, Xiao Yang Zhu

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

A high-density poly(ethylene glycol) (PEG)-coated Si(111) surface is used for the immobilization of polyhistidine-tagged protein molecules. This process features a number of properties that are highly desirable for protein microarray technology: (i) minimal nonspecific protein adsorption; (ii) highly uniform surface functionality; (iii) controlled protein orientation; and (iv) highly specific immobilization reaction without the need of protein purification. The high-density PEG-coated silicon surface is obtained from the reaction of a multi-arm PEG (mPEG) molecule with a chlorine terminated Si(111) surface to give a mPEG film with thickness of 5.2 nm. Four out of the eight arms on each immobilized mPEG molecule are accessible for linking to the chelating iminodiacetic acid (IDA) groups for the binding of Cu2+ ions. The resulting cu2+-IDA-mPEG-Si(111) surface is shown to specifically bind 6x histidinetagged protein molecules, including green fluorescent protein (GFP) and sulfotransferase (ST), but otherwise retains its inertness towards nonspecific protein adsorption. We demonstrate a particular advantage of this strategy: the possibility of protein immobilization without the need of prepurification. Surface concentrations of relevant chemical species are quantitatively characterized at each reaction step by X-ray photoelectron spectroscopy (XPS). This kind of quantitative analysis is essential in tuning surface concentration and chemical environment for optimal sensitivity in probe-target interaction.

Original languageEnglish
Pages (from-to)1965-1976
Number of pages12
JournalProteomics
Volume4
Issue number7
DOIs
Publication statusPublished - 2004 Jul

Keywords

  • Cu-IDA
  • High density PEG immobilization
  • Inert surface
  • Protein microarray

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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