Optimization of DNA-directed immobilization on mixed oligo(ethylene glycol) monolayers for immunodetection

Wen Pin Hu, Li Ya Huang, Tai Chih Kuo, Wei Wen Hu, Yung Chang, Chien-Sheng Chen, Hong Cheng Chen, Wen Yih Chen

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The development of protein chips has suffered from problems regarding long-term protein stability and activity. We present a protein sensor surface for immunodetection that is prepared by a DNA-directed protein immobilization method on a mixed self-assembled monolayer (SAM). By this approach, an immobilized single-stranded DNA (ssDNA) surface can be transferred/modified into a protein chip by flowing in ssDNA-conjugated protein when the protein chip measurement is needed. Therefore, the long-term stability of the protein chip will not be a problem for various applications. We tried various compositions for the SAM layer, the length of the ssDNA spacer, the end-point nucleotide composition, and the processes of ssDNA immobilization of the SAM for an optimized condition for shifting the DNA chip to a protein chip. The evaluations were made by using surface plasmon resonance. Our results indicated that a 50:1 ratio of oligo(ethylene glycol) (OEG)/COOH-terminated OEG and DNA sequences with 20 mer are the best conditions found here for making a protein chip via a DNA-directed immobilization (DDI) method. The designed end-point nucleotide composition contains a few guanines or cytosines, and ssDNA immobilization of the SAM by dehybridizing immobilized double-stranded DNA (dsDNA) can improve the hybridization efficiency.

Original languageEnglish
Pages (from-to)26-35
Number of pages10
JournalAnalytical Biochemistry
Volume423
Issue number1
DOIs
Publication statusPublished - 2012 Apr 1

Fingerprint

Protein Array Analysis
Ethylene Glycol
Immobilization
Monolayers
Single-Stranded DNA
Self assembled monolayers
Proteins
Nucleotides
Surface Plasmon Resonance
Protein Stability
Cytosine
Guanine
Chemical analysis
Membrane Proteins
Surface plasmon resonance

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Hu, Wen Pin ; Huang, Li Ya ; Kuo, Tai Chih ; Hu, Wei Wen ; Chang, Yung ; Chen, Chien-Sheng ; Chen, Hong Cheng ; Chen, Wen Yih. / Optimization of DNA-directed immobilization on mixed oligo(ethylene glycol) monolayers for immunodetection. In: Analytical Biochemistry. 2012 ; Vol. 423, No. 1. pp. 26-35.
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Optimization of DNA-directed immobilization on mixed oligo(ethylene glycol) monolayers for immunodetection. / Hu, Wen Pin; Huang, Li Ya; Kuo, Tai Chih; Hu, Wei Wen; Chang, Yung; Chen, Chien-Sheng; Chen, Hong Cheng; Chen, Wen Yih.

In: Analytical Biochemistry, Vol. 423, No. 1, 01.04.2012, p. 26-35.

Research output: Contribution to journalArticle

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