Quartz crystal microbalance biosensor for recombinant human interferon-β detection based on antisense peptide approach
Quartz crystal microbalance biosensor for recombinant human interferon-β detection based on antisense peptide approach
复制标题
DOI：
10.1016/j.aca.2007.03.022
复制
发表时间：
2007-05-02
影响因子：
6.2
通讯作者：
Zhao, Rui
中科院分区：
文献类型：
--
作者：
Luo, Jia;Zhang, Qundan;Zhao, Rui
利用粘附在 QCM 金表面的反义肽构建了重组人干扰素 β (rhIFN-β) 的石英晶体微天平 (QCM) 生物传感器。本研究使用了两种反义肽，均对应于 rhIFN-β 的 N 末端片段 1-14。反义肽AS-1是原始反义肽，AS-2是基于反义肽简并性的修饰反义肽。两种反义肽分别通过自组装单层 1,2-乙二硫醇固定在压电晶体的金电极上。使用石英晶体微天平流动注射分析（QCM-FIA）系统评估溶液中rhIFN-β和每种固定反义肽之间的结合亲和力。 rhIFN-β在反义肽AS-1和AS-2生物传感器上的解离常数分别为(1.89±0.101)×10(-4)和(1.22±0.0479)×10(-5)mol·L-1。结果表明AS-2对rhIFN-β的结合亲和力高于AS-1。使用每个生物传感器对 rhIFN-β 的检测是精确且可重复的。 rhIFN-β与两种生物传感器结合的线性响应范围相同，浓度范围为0.12-0.96 mg·mL(-1)。结果证明利用反义肽方法成功构建了高选择性QCM生物传感器，并证实了通过适当的序列修饰提高反义肽结合亲和力的可行性。 (c) 2007 Elsevier B.V. 保留所有权利。
Quartz crystal microbalance (QCM) biosensors for recombinant human interferon-beta (rhIFN-beta) were constructed by utilizing antisense peptides adhering to the QCM gold surfaces. Two antisense peptides, both corresponding to the N-terminal fragment 1-14 of rhIFN-beta, were used in this study. Antisense peptide AS-1 was the original antisense peptide and AS-2 was the modified antisense peptide based on the antisense peptide degeneracy. Both antisense peptides were immobilized on the gold electrodes of piezoelectric crystals, respectively, via a self-assembling monolayer of 1,2-ethanedithiol. The binding affinity between rhIFN-beta and each immobilized antisense peptide in solution was evaluated using a quartz crystal microbalance-flow injection analysis (QCM-FIA) system. The dissociation constant of rhIFN-beta on the antisense peptide AS-1 and AS-2 biosensor was (1.89 +/- 0.101) X 10(-4) and (1.22 +/- 0.0479) x 10(-5) mol L-1, respectively. The results suggested that AS-2 had a higher binding affinity to rhIFN-beta than AS-1. The detection for rhIFN-beta using each biosensor was precise and reproducible. The linear response ranges of rhIFN-beta binding to both biosensors were same with a concentration range of 0.12-0.96 mg mL(-1). The results demonstrated the successful construction of highly selective QCM biosensors using antisense peptide approach, and also confirmed the feasibility of increasing antisense peptide binding affinity by appropriate sequence modification. (c) 2007 Elsevier B.V. All rights reserved.