CAJ | 학술논문

研制基于甲胎蛋白抗体(Ab-AFP)和巯基丁二酰胺铜(Ⅱ)(CuL)共固定修饰金电极(Au| Ab-AFP/Al_2O_3/CuL)的免疫传感器,用于测定人血清中AFP抗原水平.该免疫传感器是利用自组装和溶胶-凝胶技术,将AFP抗体分子固定在CuL自组装修饰金电极表面制备而成.电极表面的CuL具有电活性,对H_2O_2有良好的电化学还原催化.当该免疫传感器在含AFP样品的溶液中于28 ℃温育30 min后,AFP抗原与Ab-AFP抗体分子的免疫结合物导致CuL的电子传递被部分阻碍,使CuL对H_2O_2 电催化还原的效率降低,电流值的下降量和AFP浓度成正比,可用于AFP定量测定.在 pH=6.5的PBS中, 28 ℃温育30 min的优化条件下,该传感器对AFP的检测线性范围为0.6～45 ng/mL,检出下限为0.088 ng/mL.该免疫传感器制备过程比基于酶标抗体修饰传感器简单,尤其是对H_2O_2催化响应迅速,稳定性好,可以在磷酸盐缓冲溶液中稳定存放25天.对于构建无试剂的免疫传感器和临床免疫分析提供了一种新颖的方法.
연제기우갑태단백항체(Ab-AFP)화구기정이선알동(Ⅱ)(CuL)공고정수식금전겁(Au| Ab-AFP/Al_2O_3/CuL)적면역전감기,용우측정인혈청중AFP항원수평.해면역전감기시이용자조장화용효-응효기술,장AFP항체분자고정재CuL자조장수식금전겁표면제비이성.전겁표면적CuL구유전활성,대H_2O_2유량호적전화학환원최화.당해면역전감기재함AFP양품적용액중우28 ℃온육30 min후,AFP항원여Ab-AFP항체분자적면역결합물도치CuL적전자전체피부분조애,사CuL대H_2O_2 전최화환원적효솔강저,전류치적하강량화AFP농도성정비,가용우AFP정량측정.재 pH=6.5적PBS중, 28 ℃온육30 min적우화조건하,해전감기대AFP적검측선성범위위0.6～45 ng/mL,검출하한위0.088 ng/mL.해면역전감기제비과정비기우매표항체수식전감기간단,우기시대H_2O_2최화향응신속,은정성호,가이재린산염완충용액중은정존방25천.대우구건무시제적면역전감기화림상면역분석제공료일충신영적방법.
A novel immunosensor for rapid separation-free determination of α-fetoprotein antigen (AFP) in human serum is proposed. The immunosensor is prepared by co-immobilizing AFP antibody(Ab-AFP) and mercapto succinic amide copper(Ⅱ)(CuL) on a glassy carbon electrode (Au) through alumina sol-gel film (Au|Ab-AFP/Al_2O_3/CuL). The electrochemical behavior of the immobilized CuL displays a surface-controlled electrode process. CuL can be used as an electro-reductive catalyzation for H_2O_2. After the immunosensor is incubated with AFP solution at 28 ℃ for 30 min, the access of activity center of CuL is partly inhibited, which leads to a linear decrease of the catalytic efficiency of CuL by H_2O_2 in two AFP concentration ranges from 0.6 to 45 ng/mL. Under optimal conditions the detection limit for AFP immunoassay is 0.088 ng/mL(3σ). The immunosensor shows a good accuracy and acceptable storage stability, precision and reproducibility. This method decreases the detection cost and shortens the analytical time, thus would be valuable for clinical immunoassay.