Comparison of DNA, aminoethylglycyl PNA and pyrrolidinyl PNA as probes for detection of DNA hybridization using surface plasmon resonance technique
Comparison of DNA, aminoethylglycyl PNA and pyrrolidinyl PNA as probes for detection of DNA hybridization using surface plasmon resonance technique
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DOI：
10.1016/j.bios.2009.09.028
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发表时间：
2010-01-15
影响因子：
12.6
通讯作者：
Su, Xiaodi
中科院分区：
文献类型：
--
作者：
Ananthanawat, Cheeraporn;Vilaivan, Tirayut;Su, Xiaodi
以D-脯氨酰-2-氨基环戊烷羧酸为骨架的吡咯烷肽核酸（acpcPNA）是一种新型的DNA杂交检测探针。在这项研究中，生物素化acpcPNA固定在表面等离子体共振（SPR）传感器芯片通过生物素-链霉亲和素相互作用的固相DNA杂交。本文用SPR技术对acpcPNA、DNA和相同序列的常规肽核酸（aegPNA）探针在各个重要方面进行了比较。这些包括离子强度对杂交效率的影响，检测靶DNA中错配的特异性，与靶DNA结合的方向（平行或反平行），以及靶DNA浓度对杂交效率的影响。结果表明，固定化acpcPNA探针与aegPNA（和/或DNA）探针具有明显的杂交特性。包括更高的单碱基错配灵敏度、反平行选择性和靶杂交的低离子强度依赖性。这些性质证实了acpcPNA作为传感器探针用于临床和诊断应用的适用性。适当选择再生条件（10 mM NaOH. 2分钟暴露），传感器可以重复用于多个杂交循环，每个再生循环的杂交活性损失少至1.3%。(C)2009爱思唯尔有限公司版权所有。
Pyrrolidinyl peptide nucleic acid bearing a D-prolyl-2-aminocyclopentanecarboxylic acid backbone (acpcPNA) has been evaluated as a new sensing probe for detection of DNA hybridization. In this study, the biotinylated acpcPNA was immobilized on surface plasmon resonance (SPR) sensor chips via biotin-streptavidin interactions for solid-phase DNA hybridization. A critical comparison between acpcPNA, DNA and conventional peptide nucleic acid (aegPNA) probes of the same sequence was made by means of SPR on various important aspects. These include the effect of ionic strength on hybridization efficiency, the specificity to detect the mismatch(es) in target DNAs, the direction of binding (parallel or antiparallel) to target DNAs, and the effect of target DNA concentration on hybridization efficiency. Results indicated that the immobilized acpcPNA probe possesses distinct hybridization properties relative to aegPNA (and/or DNA) counterparts. including a higher single-base mismatch sensitivity, antiparallel selectivity and low ionic strength dependence of target hybridization. These properties substantiate the acpcPNA applicability as sensor probes for clinical and diagnostic applications. With a proper selection of regeneration conditions (10 mM NaOH. 2 min exposure), the sensor can be reused for multiple cycles of hybridization with as little as 1.3% loss in hybridization activity per regeneration cycle. (C) 2009 Elsevier B.V. All rights reserved.