Optimizing Multiple Analyte Injections in Surface Plasmon Resonance Biosensors with Analytes having Different Refractive Index Increments

Abstract

Surface plasmon resonance-based biosensors have been successfully applied to the study of the interactions between macromolecules and small molecular weight compounds. In an effort to increase the throughput of these SPR-based experiments, we have already proposed to inject multiple compounds simultaneously over the same surface. When specifically applied to small molecular weight compounds, such a strategy would however require prior knowledge of the refractive index increment of each compound in order to correctly interpret the recorded signal. An additional experiment is typically required to obtain this information. In this manuscript, we show that through the introduction of an additional global parameter corresponding to the ratio of the saturating signals associated with each molecule, the kinetic parameters could be identified with similar confidence intervals without any other experimentation.

Determination of the composition of heterogeneous binder solutions by surface plasmon resonance biosensing

“…No regeneration step was needed between injections as the dissociation was complete in every case. This agrees with previous studies that used this system [10][11][12]20,28 . The SPR biosensor used in this study (Biacore T100) uses a detection mode based on varying the incident angle while keeping the wavelength constant.…”

“…The said analyte may be purified 1 or in a crude extract [2][3][4] . Through the years, important advances were made in the development of optimized experimental protocols 5 and data analysis methods [6][7][8][9][10][11][12] . Robust data analysis software packages were created and contributed to the success of this label-free technology [13][14][15][16] .…”

“…Recent efforts have brought the SPR technology at the forefront of drug development for (i) the off-line 17 or at-line 18 characterization of the critical quality attributes of biotherapeutics or (ii) for the screening of large libraries of drug candidates 19 . On that specific note, our group recently demonstrated that the throughput of a SPR-based experiment could be improved by injecting two analytes simultaneously, since the binding kinetics of both analytes could be identified in only one set of experiments 10,12 . These methods have yet to be expanded to the study of more than two analytes.…”

“…In a second part, we exploit the prior knowledge of the kinetic parameters of a set of analytes to estimate the composition of a mixture of these analytes by performing a sensorgram analysis. We validated our algorithms using a well-characterized model system [10][11][12]20,28 ,…”

Temporal response of biochemical and biological sensors with bimodal surface adsorption from a finite sample

“…To analyze the measurement results a simple linear mathematical model of sensor temporal response is commonly applied (Mehand et al 2015 ), in which it is assumed that only adsorption–desorption (AD) process of analyte particles on the sensing surface influences the sensor response. Alternatively, a somewhat more comprehensive model is used that takes into account the coupling of AD process and mass transfer in the sensor microfluidic chamber (Myszka et al 1998 ; Anderson et al 2011 ).…”