Faculty of Medicine, the School of Pharmacy Institute for Drug Research, The Hebrew University of Jerusalem, P.O.Box 12065, Jerusalem, 91120, Israel.
Pharm Res. 2012 Apr;29(4):983-93. doi: 10.1007/s11095-011-0638-8. Epub 2011 Dec 10.
A real time detection of gastric cancer-associated biomarker molecules in the lumen of the stomach could assist in early detection of this multi-step malignancy.
Employing α1-antitrypsin precursor (A1AT) as a secreted biomarker model, a platform with immunoassay capabilities, comprising sensing and detecting compartments was developed. It was made of a microarray-type functionalized glass, containing a high density of amine groups. Trypsin, the capturing moiety, was immobilized to the glass surface with the aid of a PEG-based spacer mixture, identified as being crucial for both capturing and detecting properties. The detecting compartment contained near infrared fluorescently labeled nanoparticles conjugated to A1AT-specific antibodies, aimed at generating an optical signal, detectable by a conventional endoscope or a video capsule.
The specific recognition reaction between the captured A1AT and the immuno-nanoparticles generated a profound fluorescence with a signal to noise ratio (SNR) of 12-32, in a biomarker-concentration dependent manner. Moreover, the optical recognition signal was intense enough to be detected by a video capsule simulator (with optical detection capabilities of a video capsule) with a SNR of 6-20.
This platform could serve as a real time diagnostic kit for early detection of a secreted biomarker of gastric cancer.
实时检测胃腔中与胃癌相关的生物标志物分子,有助于早期发现这种多步骤发生的恶性肿瘤。
采用α1-抗胰蛋白酶前体(A1AT)作为分泌性生物标志物模型,开发了一种具有免疫分析功能的平台,包括传感和检测两个部分。该平台由微阵列型功能化玻璃组成,玻璃表面含有高密度的胺基。用基于聚乙二醇的间隔混合剂将捕获部分胰蛋白酶固定在玻璃表面上,这种间隔混合剂对于捕获和检测性能都非常重要。检测部分含有近红外荧光标记的纳米粒子,与 A1AT 特异性抗体偶联,旨在产生光学信号,可通过传统内窥镜或视频胶囊进行检测。
在生物标志物浓度依赖的情况下,捕获的 A1AT 与免疫纳米粒子之间的特异性识别反应产生了强烈的荧光,信噪比(SNR)为 12-32。此外,光学识别信号足够强,可以通过视频胶囊模拟器(具有视频胶囊的光学检测能力)检测到,信噪比为 6-20。
该平台可作为实时诊断试剂盒,用于早期检测胃癌的分泌性生物标志物。