Expanding the ProteOn XPR36 biosensor into a 36-ligand array expedites protein interaction analysis

Area of Science:

• Biochemistry
• Immunology
• Analytical Chemistry

Background:

• Biosensor technology is crucial for characterizing molecular interactions.
• High-throughput analysis is needed for comprehensive kinetic and epitope studies.
• The ProteOn XPR36 biosensor offers potential for multiplexed assays.

Purpose of the Study:

• To demonstrate methods for expanding the throughput of the ProteOn XPR36 biosensor.
• To enable simultaneous kinetic characterization of multiplexed antibody formats.
• To facilitate detailed epitope binning and mapping studies.

Main Methods:

• Utilizing the ProteOn XPR36 biosensor in multiplexed formats for kinetic characterization.
• Performing pairwise epitope binning using the ProteOn 36-ligand array.
• Conducting epitope mapping studies with biotinylated peptides.
• Comparing results with Biacore 2000 and Octet QK384 biosensor platforms.

Main Results:

• Kinetic rate constants determined on the ProteOn XPR36 correlated with Biacore 2000.
• ProteOn GLC chip data showed closer agreement with Biacore C1 chip than CM4 chip.
• Epitope binning data from ProteOn and Octet QK384 were similar.
• All three platforms effectively identified antibodies binding to linear epitopes.

Conclusions:

• The ProteOn XPR36 biosensor can be utilized in alternative formats for significantly higher throughput.
• The demonstrated methods allow for simultaneous kinetic characterization and epitope analysis.
• The ProteOn XPR36 provides accurate and comparable results to established biosensor platforms.