摘要
目的探索适配子压电石英芯片最佳再生试剂,对固定好的适配子型芯片保存性能进行检测。方法用生物素-亲和素法将针对人IgE的适配子探针固定在压电传感器的金膜表面,分别用HCl、NaOH、EDTA、尿素、甲酰胺5种试剂对适配子芯片进行再生实验,选取较好再生试剂;将固定好的适配子芯片保存在PBS缓冲液(0.1%叠氮钠)中,对芯片的保存时间进行测定。结果用EDTA对适配子芯片进行再生效果最好,重复检测5次后芯片响应信号仍在第1次检测的>90%,此外HCl也可对适配子芯片重复利用3次;固定好的适配子芯片在加入防腐剂的结合缓冲液中可长期保存,保存21d后信号响应无明显下降。结论与抗体压电传感器相比较,适配子压电传感器有一定优势,适配子芯片成本更低,再生能力强,固定好的芯片保存时间长,体现出适配子检测的优越性。
OBJECTIVE To study the optimal reagent for regeneration of aptamer-eoated piezoelectric quartz crystal chips and detect storage ability of aptamer-eoated chips. METHODS Anti-IgE aptamers were immobilized on the gold surface of piezoelectric quartz crystal biosensor with biotin-avidin method. The reaction between anti-IgE aptamers and IgE was monitored in time on piezoelectric sensor. We tried to regenerate the sensor surface after binding IgE by rinsing the aptamer-eoated chips with HCI, NaOH, EDTA, urea and formamide individually. Aptamer-eoated chips were stored in PBS with 0.1% sodium azide and the stored chips were used to detect IgE in 35 days. RESULTS Of the five reagents, EDTA was the best one for regeneration of aptamer-coated chips and the sensor could retain 91.5% of the original detecting signals after five regeneration cycles. Moreover, the aptamer- coated chips could be stored in the binding buffer for 21 days without obvious loss of activity. CONCLUSIONS Compared with antibody-based piezoelectric sensor, aptamer-based sensor has lower cost, more regeneration cycles and longer time for the storage of chips. This series of experiments shows the superiority of aptamer detection.
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2008年第3期356-358,共3页
Chinese Journal of Nosocomiology
基金
国家自然科学基金资助项目(30400420)
