School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022, PR China.
Biomaterials. 2011 Mar;32(8):2117-23. doi: 10.1016/j.biomaterials.2010.11.058. Epub 2010 Dec 24.
BRCAl is an anti-oncogene in women, who are genetically predisposed to breast and ovary cancer. The detection of BRCA1 can offer an opportunity to characterize the function of genetic features in breast and ovarian cancer and to screen breast or ovarian cancer patients. In this study, we designed a new label and fabricated a novel sandwich-type electrochemical immunoassay for the ultrasensitive detection of BRCAl. Horseradish peroxidase (HRP) was entrapped in the pores of amino-group functionalized SBA-15 and the secondary antibody (Ab₂) combined with SBA-15 by covalent bond. Ionic liquid (IL) was added into the mixed solution of SBA-15/HRP/Ab₂ and application of IL increased the electrochemical activity of HRP and promoted electron transport. The synergistic effect between IL, SBA-15, Ab₂ and HRP could retain the bioactivity of HRP and Ab₂. The sensitivity of the sandwich-type immunosensor using SBA-15/HRP/Ab₂/BMIM·BF₄ as labels for BRCA1 detection was much higher than that using either SBA-15/HRP/Ab₂ or SBA-15/Ab₂ as labels. Under optimal conditions, the electrochemical immunoassay exhibited a wide working range from 0.01 to 15 ng/mL with a detection limit of 4.86 pg/mL BRCA1. The precision, reproducibility, and stability of the immunoassay were acceptable.
BRCA1 是女性的一种抑癌基因，她们具有乳腺癌和卵巢癌的遗传易感性。BRCA1 的检测为研究乳腺癌和卵巢癌的遗传特征的功能以及筛查乳腺癌或卵巢癌患者提供了机会。本研究设计了一种新的标签，并构建了一种新型三明治型电化学免疫分析方法，用于超灵敏检测 BRCAl。辣根过氧化物酶（HRP）被包埋在氨基功能化 SBA-15 的孔中，并且第二抗体（Ab₂）通过共价键与 SBA-15 结合。将离子液体（IL）加入到 SBA-15/HRP/Ab₂ 的混合溶液中，IL 的加入增加了 HRP 的电化学活性并促进了电子传递。IL、SBA-15、Ab₂ 和 HRP 之间的协同作用可以保留 HRP 和 Ab₂ 的生物活性。使用 SBA-15/HRP/Ab₂/BMIM·BF₄ 作为 BRCA1 检测的标签的三明治型免疫传感器的灵敏度远高于单独使用 SBA-15/HRP/Ab₂ 或 SBA-15/Ab₂ 作为标签的灵敏度。在最佳条件下，电化学免疫分析的工作范围从 0.01 到 15 ng/mL，检测限为 4.86 pg/mL BRCA1。免疫分析的精密度、重现性和稳定性均良好。
