A Boronate Affinity-Assisted SERS Tag Equipped with a Sandwich System for Detection of Glycated Hemoglobin in the Hemolysate of Human Erythrocytes
详细信息 查看全文
- 作者：Duygu Deniz Usta ; Kouroush Salimi ; Asli Pinar ; İlknur Coban ; Turgay Tekinay ; Ali Tuncel
- 刊名：ACS Applied Materials & Interfaces
- 出版年：2016
- 出版时间：May 18, 2016
- 年：2016
- 卷：8
- 期：19
- 页码：11934-11944
- 全文大小：595K
- 年卷期：0
- ISSN：1944-8252
文摘
Phenylboronic acid-functionalized, Ag shell-coated, magnetic, monodisperse polymethacrylate microspheres equipped with a glycoprotein-sensitive sandwich system were proposed as a surface-enhanced Raman scattering (SERS) substrate for quantitative determination of glycated hemoglobin (HbA1c). The magnetization of the SERS tag and the formation of the Ag shell on the magnetic support were achieved using the bifunctional reactivity of newly synthesized polymethacrylate microspheres. The hemolysate of human red blood cells containing both HbA1c and nonglycated hemoglobin was used for determination of HbA1c. The working principle of the proposed SERS tag is based on the immobilization of HbA1c by cyclic boronate ester formation between glycosyl residues of HbA1c and boronic acid groups of magnetic polymethacrylate microspheres and the binding of p-aminothiophenol (PATP)-functionalized Ag nanoparticles (Ag NPs) carrying another boronic acid ligand via cyclic boronate ester formation via unused glycosyl groups of bound HbA1c. Then, in situ formation of a Raman reporter, 4,4′-dimercaptoazobenzene from PATP under 785 nm laser irradiation allowed for the quantification of HbA1c bound onto the magnetic SERS tag, which was proportional to the HbA1c concentration in the hemolysate of human erythrocytes. The sandwich system provided a significant enhancement in the SERS signal intensity due to the plasmon coupling between Ag NPs and Ag shell-coated magnetic microspheres, and low HbA1c concentrations down to 50 ng/mL could be detected. The calibration curve obtained with a high correlation coefficient between the SERS signal intensity and HbA1c level showed the usability of the SERS protocol for the determination of the HbA1c level in any person.