Abstract: Glycoproteins are a class of proteins with significant biological functions and clinical implications. Due to glycoproteins' reliability for the quantitative analysis, they have been used as biomarkers and therapeutic targets for disease diagnosis. We propose a sandwich structure-based boronate affinity biosensor that can separate and detect target glycoproteins by magnetic separation and Surface-enhanced Raman scattering (SERS) probes. The biosensor relies on boronic acid affinity magnetic molecularly imprinted polymer (MMIPs) with pH response as "capturing probe" for glycoproteins, and Au-MPBA@Ag modified with 4-mercaptophenylboronic acid (MPBA) as SERS probes, among which, MPBA has both strong SERS activity and can specifically recognize and bind to glycoproteins. MMIPs ensured specific and rapid analysis, and SERS detection provided high sensitivity. The proposed boronate affinity SERS strategy exhibited universal applicability and provided high sensitivity with limit of detection of 0.053 ng/mL and 0.078 ng/mL for horseradish peroxidase and acid phosphatase, respectively. Ultimately, the boronate affinity SERS strategy was successfully applied in detection of glycoprotein in spiked serum sample with recovery between 90.6% and 103.4%, respectively. In addition, this study used a portable Raman meter, which can meet the requirements of point-of-care testing. The biosensor presented here also has advantages in terms of cost-effectiveness, stability, and detection speed
Author keywords: boronate affinity, SERS, magnetic molecularly imprinted polymer, Glycoprotein, Controllable-oriented surface imprinting