摘要
目的提高检测B型葡萄球菌肠毒素的Love波免疫传感器的再生能力,以优化Love波免疫传感器的设计。方法传感器由压电石英基片、叉指换能器和SiO2声波导层构成。采用戊二醛共价交联法构建抗体分子探针传感界面。为提高传感器的再生能力,选用NaBH4还原连接于戊二醛和抗体分子之间的Schiff’s base结构,通过实验确定NaBH4还原步骤和最佳浓度,并用原子力显微镜(AFM)对改良后的戊二醛法固定抗体以及捕获SEB后的表面进行成像表征。结果采用优化后的固定化方案制备的免疫传感器对10-6g.mL-1SEB有明显响应,并可以重复使用3次,NaBH4的最佳浓度为100 mmol.L-1。结论先共价交联抗体分子,再用浓度为100 mmol.L-1的NaBH4还原共价双键为稳定的单键能明显改善Love传感器对SEB的响应能力和再生能力。
Objective To improve the regeneration of a Love mode surface acoustic wave （SAW） immunosensor for staphylococcal enterotoxin B （SEB） detection, so as to optimize the design of the Love wave Methods The Love wave device with the substrate of ST-cut quartz, IDTs and the guiding layers of SiO2 was fabricated. In the immobilization of antibodies to the sensor by glutaraldehyde cross-linking procedure, a selective reducing gent, sodium borohydride （NaBH4）, with different concentration and manner was used to reduce the Schiff＇s base resulting from the reaction of glutaraldehyde-activated surface with antibody amino groups to improve the regeneration of the immunosensor. The surface morphology of the active area was characterized by using atomic force microscopy. Results Such a preparation could greatly increase regeneration and sensitivity of the immunosensor, up to 10.6 g·mL^-1 SEB for 3 successive assay repetitions without significant loss of sensitivity. The optimized concentration of NaBH4 was 100 mmol·L^-1. Conclusion The immobilization of antibody on surface immediately followed by reduction with 100 mmol·L^-1 NaBH4 is the best approach. Glutaraldehyde-NaBH4 procedure can allow Love wave immunosensor with better SEB-binding capabilities and better regeneration.
出处
《解放军预防医学杂志》
CAS
北大核心
2006年第4期250-253,共4页
Journal of Preventive Medicine of Chinese People's Liberation Army
基金
国家自然科学基金资助项目课题(No.30371218)
