Abstract：
This work describes the development of a label-free (LF) biosensing platform for the direct detection of targets based on diffractive structures fabricated with acrylamide-based hydrogels biofunctionalized with proteins and antibodies. Hydrogels containing Bovine Serum Albumin protein (BSA) with different crosslinking degrees were synthesized and characterized to find the optimal conditions for the suitable fabrication of surface relief gratings (SRGs). The bioavailability of BSA-functionalized hydrogels for the specific recognition of anti-BSA antibodies was verified by fluorescence detection. After the hydrogel-based SRG fabrication, diffraction efficiency measures at two different laser wavelengths were used for the direct LF detection of anti-BSA antibodies. The limit of detection in the sub mg L-1 range was read. Additionally, SRGs were prepared with hydrogels biofunctionalized with anti-rabbit antibodies for the direct detection of IgGs from rabbit serum, obtaining similar analytical performance without the necessity of labeling or applying amplification strategies.
摘要：
这项工作描述了无标记（LF）生物传感平台的开发，该平台用于基于用基于丙烯酰胺的水凝胶用蛋白质和抗体生物功能化的衍射结构直接检测目标。合成了含有不同交联度的牛血清白蛋白（BSA）的水凝胶，并对其进行了表征，以找到合适的表面浮雕光栅（SRG）制造的最佳条件。通过荧光检测验证了BSA官能化水凝胶用于特异性识别抗BSA抗体的生物利用度。在基于水凝胶的SRG制造之后，在两个不同激光波长下的衍射效率测量用于抗BSA抗体的直接LF检测。读取亚mgL-1范围内的检测极限。此外,用抗兔抗体生物功能化的水凝胶制备SRG，用于直接检测兔血清中的IgG，获得相似的分析性能，而无需标记或应用扩增策略。
