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

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Title:	Optimization of DNA-directed immobilization on mixed oligo(ethylene glycol) monolayers for immunodetection
Authors:	陳文逸;Hu, Wen-Pin;Huang, Li-Ya;Kuo, Tai-Chih;Hu, Wei-Wen;Chang, Yung;Chen, Chien-Sheng;Chen, Hong-Cheng;Chen, Wen-Yih
Contributors:	工學院化學工程與材料工程學系
Keywords:	Antibodies - chemistry;Antibodies - immunology;Base Sequence;DNA microarrays;DNA, Single-Stranded - chemistry;DNA, Single-Stranded - metabolism;DNA-directed immobilization (DDI);ethylene glycol;Ethylene Glycol - chemistry;hybridization;immobilization;Immunoassay - methods;Muramidase - analysis;Muramidase - immunology;nucleotide sequences;Oligo(ethylene glycol) (OEG);Optimization;Protein Array Analysis;Proteins - analysis;Proteins - immunology;Self-assembled monolayer (SAM);Serum Albumin - analysis;Serum Albumin - immunology;single-stranded DNA;Surface Plasmon Resonance;Surface plasmon resonance (SPR)
Date:	2012-04-01
Issue Date:	2026-04-21 13:53:49 (UTC+8)
Publisher:	Academic Press Inc.;United States: Elsevier Inc
Abstract:	摘要： 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 20mer 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. 其他題名： Anal Biochem 出版者： United States: Elsevier Inc 出版日期： 2012-04-01 出處： Analytical biochemistry, 2012-04, Vol.423 (1), p.26-35 版權： 2012 Elsevier Inc. 版權： Copyright Â© 2012 Elsevier Inc. All rights reserved. 識別號： ISSN: 0003-2697 識別號： ISSN: 1096-0309 識別號： EISSN: 1096-0309 識別號： DOI: 10.1016/j.ab.2011.12.043 識別號： PMID: 22285698
Appears in Collections:	[Department of Chemical and Materials Engineering] journal & Dissertation

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