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

doi:10.1016/j.ab.2011.12.043

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

Department of Food Safety / Hygiene and Risk Management

Research output: Contribution to journal › Article › peer-review

16 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 language	English
Pages (from-to)	26-35
Number of pages	10
Journal	Analytical Biochemistry
Volume	423
Issue number	1
DOIs	https://doi.org/10.1016/j.ab.2011.12.043
Publication status	Published - 2012 Apr 1

All Science Journal Classification (ASJC) codes

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.ab.2011.12.043

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

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.",

author = "Hu, \{Wen Pin\} and Huang, \{Li Ya\} and Kuo, \{Tai Chih\} and Hu, \{Wei Wen\} and Yung Chang and Chen, \{Chien Sheng\} and Chen, \{Hong Cheng\} and Chen, \{Wen Yih\}",

note = "Funding Information: The authors gratefully acknowledge the financial support provided by the National Science Council, Taiwan , for the Center for Dynamical Biomarkers and Translational Medicine, National Central University, Taiwan ( NSC 99-2911-I-008-100 and NSC 100-2320-B-468-001 ).",

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AU - Hu, Wen Pin

AU - Huang, Li Ya

AU - Kuo, Tai Chih

AU - Hu, Wei Wen

AU - Chang, Yung

AU - Chen, Chien Sheng

AU - Chen, Hong Cheng

AU - Chen, Wen Yih

N1 - Funding Information: The authors gratefully acknowledge the financial support provided by the National Science Council, Taiwan , for the Center for Dynamical Biomarkers and Translational Medicine, National Central University, Taiwan ( NSC 99-2911-I-008-100 and NSC 100-2320-B-468-001 ).

PY - 2012/4/1

Y1 - 2012/4/1

N2 - 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.

AB - 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.

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

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