新型血清CYFRA21-1检测电化学生物传感器的研制

A novel electrochemical biosensor for the detection of serum CYFRA21-1

目的构建一种用于检测细胞角蛋白19片段(CYFRA21-1)的高灵敏电化学生物传感器。方法合成三维石墨烯@金纳米粒子(3D-G@Au)复合材料,用其修饰玻碳电极,通过壳聚糖、戊二醛与抗CYFRA21-1抗体交联,构建用于检测CYFRA21-1的3D-G@Au的电化学生物传感器。对构建成功的3D-G@Au的电化学生物传感器的电化学性能及检测性能进行初步评价。结果循环伏安图显示裸玻碳电极、3D-G修饰玻碳电极和3D-G@Au修饰玻碳电极的氧化还原峰逐渐升高,提示3D-G@Au电化学生物传感器构建成功。传感器抗原-抗体相互作用的最佳温育时间为60 min,最佳孵育温度为35℃。采用3D-G@Au电化学生物传感器检测CYFRA21-1,线性范围为0.1~300.0 ng/mL,检测限为0.1 ng/mL(信噪比=3),10 mmol/L维生素C、10mmol/L多巴胺和10mmol/L尿酸对检测无干扰,重现性较好[相对标准偏差(RSD)分别为1.08%、3.11%],与化学发光法比较,RSD为1.96%~4.78%。制备好的3D-G@Au电化学生物传感器可在4℃条件下稳定30 d。结论构建的3D-G@Au电化学生物传感器可用于血清CYFRA21-1的检测。

Objective To construct a highly sensitive electrochemical biosensor for the detection of cytokeratin 19-fragment(CYFRA21-1).Methods Three-dimensional graphene@gold nanoparticles(3D-G@Au)nanocomposite was modified on glassy carbon electrode surface to enhance the conductivity of electrochemical biosensor.Anti-CYFRA21-1 was captured and fixed on the modified 3D-G@Au glassy carbon electrode through the cross-linking of chitosan and glutaraldehyde.The 3D-G@Au electrochemical biosensor had been completed as to determine CYFRA21-1.The electrochemical and determination performance of successfully constructed 3D-G@Au electrochemical biosensor was evaluated preliminarily.Results The cyclic voltammetry showed that the redox peaks of bare glassy carbon electrode,3D-G modified glassy carbon electrode and 3D-G@Au modified glassy carbon electrode increased gradually,indicating that the 3D-G@Au electrochemical biosensor was successfully constructed.The optimal incubation time of antigen-antibody interaction was 60 min,and the optimal incubation temperature was 35℃.CYFRA21-1 was detected by 3D-G@Au electrochemical biosensor,the linear range was 0.1-300.0 ng/mL,and the determination limit was 0.1 ng/mL(signal-to-noise ratio=3).10 mmol/L Vit C,10 mmol/L DA and 10 mmol/L UA had no interference to the determination,and the reproducibility was good[relative standard deviations(RSD)were 1.08%and 3.11%],compared with that of chemiluminescence method(RSD was 19.6%-4.78%).The 3D-G@Au electrochemical biosensor can be stable for 30 d at 4℃.Conclusions It is potential for the determination of serum CYFRA21-1 with 3D-G@Au electrochemical biosensor.