Department of Chemistry, University of South Florida, 4202 E. Fowler Ave., Tampa, FL 33620-5250, USA.
J Immunol Methods. 2012 Dec 14;386(1-2):1-9. doi: 10.1016/j.jim.2012.08.007. Epub 2012 Aug 15.
The formation of a thin antibody film on a glass surface using pneumatic spray was investigated as a potential immobilization technique for capturing pathogenic targets. Goat-Escherichia coli O157:H7 IgG films were made by pneumatic spray and compared against the avidin-biotin bridge immobilized films by assaying with green fluorescent protein (GFP) transformed E. coli O157:H7 cells and fluorescent reporter antibodies. Functionality, stability, and immobilization of the films were tested. The pneumatic spray films had lower fluorescence intensity values than the avidin-biotin bridge films but resulted in similar detection for E. coli O157:H7 at 10(5)-10(7)cells/ml sample concentrations with no detection of non-E. coli O157:H7 strains. Both methods also resulted in similar percent capture efficiencies. The results demonstrated that immobilization of antibody via pneumatic spray did not render the antibody non-functional and produced stable antibody films. The amount of time necessary for immobilization of the antibody was reduced significantly from 24h for the avidin-biotin bridge to 7 min using the pneumatic spray technique, with additional benefits of greatly reduced use of materials and chemicals. The pneumatic spray technique promises to be an alternative for the immobilization of antibodies on glass slides for capturing pathogenic targets and use in biosensor type devices.
采用气动喷雾在玻璃表面形成薄的抗体膜，作为捕获致病目标的潜在固定化技术进行了研究。通过气动喷雾制备了山羊-大肠杆菌 O157:H7 IgG 膜，并通过与绿色荧光蛋白 (GFP) 转化的大肠杆菌 O157:H7 细胞和荧光报告抗体进行检测，与亲和素-生物素桥固定化膜进行了比较。对膜的功能、稳定性和固定化进行了测试。气动喷雾膜的荧光强度值低于亲和素-生物素桥膜，但在 10(5)-10(7)细胞/ml 样品浓度下对大肠杆菌 O157:H7 的检测结果相似，对非大肠杆菌 O157:H7 菌株没有检测到。两种方法的捕获效率也相似。结果表明，通过气动喷雾固定化抗体不会使抗体失去功能，并产生稳定的抗体膜。与亲和素-生物素桥方法相比，气动喷雾技术固定抗体所需的时间从 24 小时显著减少到 7 分钟，并且大大减少了材料和化学品的使用。气动喷雾技术有望成为在玻片上固定抗体以捕获致病目标并用于生物传感器类型设备的替代方法。
