Detection of Folate Binding Protein with Enhanced Sensitivity Using a Functionalized Quartz Crystal Microbalance Sensor

Walter A. Henne, Department of Chemistry, Purdue University
Derek D. Doorneweerd, Department of Chemistry, Purdue University
Joonhyung Lee, School of Chemical Engineering, Purdue University
Philip S. Low, Department of Chemistry
Cagri Savran, Birck Nanotechnology Center and School of Mechanical Engineering, Purdue University

Date of this Version

July 2006

Citation

Analytical Chemistry 2006, 78, 4880-4884

This document has been peer-reviewed.

 

Abstract

In this report, we describe the development of a quartz crystal microbalance biosensor for detection of folate binding protein (FBP). Using a simple folate—BSA conjugate absorbed onto a Au-coated quartz sensor, a detection limit of 30 nM was achieved. Binding of FBP to the sensor surface could be blocked at concentrations as high as 1 uM with a 100-fold excess of folic acid, indicating the specificity of the folate—FBP interaction and the absence of nonspecific binding to the functionalized surface. Moreover, capture could be achieved in the presence of blood serum, making the assay amendable to the analysis of bodily fluids. Further signal enhancement based on an anti-FBP antibody and protein-A-coated gold nanosphere sandwich assay extended the detection limit to 50 pM (~3 orders-of-magnitude improvement). Given the overexpression of FBP in certain malignancies and inflammatory disorders, we expect the methodology described here to be useful to detect FBP as a possible biomarker for disease diagnosis.

 

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