Information-rich High Throughput Screening with Multiplexing Surface Plasmon Resonance

Surface Plasmon Resonance is an established and widely used biophysical technology in screening and lead development campaigns for novel drug candidates. The real-time, label-free analysis of interactions offers additional insights into kinetics unlike endpoint assays like fluorescence-based assays. Multiplexing SPR systems further allow to study the interaction of a drug candidate against multiple targets simultaneously; a task which requires multiple repetitions of the same screening with other techniques. However, the throughput is often comparably low and the number of targets in a fast, efficient quantitative screening campaign is severely limited.

We present an assay set-up that allows the binding assessment of a single analyte against dozens of targets or the affinity determination of one analyte against multiple targets with a single injection. Partnered with our established SPR^* detection system, it’s possible to perform measurements also at high molecular weight differences between analyte and target of 10^-3. Furthermore, this assay set-up facilitates the determination of thermodynamic constants at low running times due to parallel readout. Access to this information is typically relevant in subsequent lead development processes.

We tested the assay set-up with multiple established assay modes for two model cases. The interaction of oligonucleotides against complementary strands and of a set of sulphonamides against multiple carbonic anhydrase isozymes confirmed the performance and robustness. Additionally, the sensitivity of our novel assay set-up was assessed with an interaction of DNP-modified amino acids (241-283 Da) against specific antibodies (150 kDa).

We herewith present a SPR assay set-up with industry-leading throughput in affinity determination and multiplexing capabilities for all types of analytes. Thus, the extension of typical parameters from a SPR experiment by a quantitative selectivity assessment allows for a more informed hit selection process in screening campaigns.