Significance Testing and Multivariate Analysis of Datasets from Surface Plasmon Resonance and Surface Acoustic Wave Biosensors: Prediction and Assay Validation for Surface Binding of Large Analytes

Abstract

In this study, we performed uni- and multivariate data analysis on the extended binding curves of several affinity pairs: immobilized acetylcholinesterase (AChE)/bioconjugates of aflatoxin B1(AFB1) and immobilized anti-AFB1 monoclonal antibody/AFB1-protein carriers. The binding curves were recorded on three mass sensitive cells operating in batch configurations: one commercial surface plasmon resonance (SPR) sensor and two custom-made Love wave surface-acoustic wave (LW-SAW) sensors. We obtained 3D plots depicting the time-evolution of the sensor response as a function of analyte concentration using real-time SPR binding sensograms. These “calibration” surfaces exploited the transient periods of the extended kinetic curves, prior to equilibrium, creating a “fingerprint” for each analyte, in considerably shortened time frames compared to the conventional 2D calibration plots. The custom-made SAW sensors operating in different experimental conditions allowed the detection of AFB1-protein carrier in the nanomolar range. Subsequent statistical significance tests were performed on unpaired data sets to validate the custom-made LW-SAW sensors.

Photoluminescence label-free immunosensor for the detection of Aflatoxin B1 using polyacrylonitrile/zinc oxide nanofibers

“…Due to the small size of AFB1 molecules, it can be even undetectable by the Surface Plasmon Resonance based approach (SPR), which is considered to be the most sensitive label-free techniques. In order to solve this problem the "weighting agents" such as bovine serum albumin (BSA) has been used [10]. However, BSA weighting agents can adversely affect detection selectivity; therefore, the use of highly selective receptors (for example, nanobodies against AFB1) is preferred over the AFB1-BSA using [11].…”