Changes in mass loading on the surface of acoustic biosensors result in output frequency shifts which provide precise measurements of analytes. Therefore, to detect a particular biomarker, the sensor delay path must be judiciously designed to maximize sensitivity and specificity. B-cell lymphoma 2 protein (Bcl-2) found in urine is under investigation as a biomarker for non-invasive early detection of ovarian cancer. In this study, surface chemistry and biofunctionalization approaches were evaluated for their effectiveness in presenting antibodies for Bcl-2 capture while minimizing non-specific protein adsorption. The optimal combination of sequentially adsorbing protein A/G, anti-Bcl-2 IgG and Pluronic F127 onto a hydrophobic surface provided the greatest signal-to-noise ratio and enabled the reliable detection of Bcl-2 concentrations below that previously identified for early stage ovarian cancer as characterized by a modified ELISA method. Finally, the optimal surface modification was applied to a prototype acoustic device and the frequency shift for a range of Bcl-2 concentration was quantified to demonstrate the effectiveness in surface acoustic wave (SAW)-based detection applications. The surface functionalization approaches demonstrated here to specifically and sensitively detect Bcl-2 in a working ultrasonic MEMS biosensor prototype can easily be modified to detect additional biomarkers and enhance other acoustic biosensors.
Keywords: Bcl-2; bioconjugation; early detection; microelectromechanical systems (MEMS); ovarian cancer; point-of-care; polyethylene glycol (PEG); self-assembled monolayer (SAM); sensor; surface acoustic wave (SAW).