title: Expanding the ProteOn XPR36 biosensor into a 36-ligand array expedites protein interaction analysis.
abstract: Here we demonstrate methods to expand the throughput of the ProteOn XPR36 biosensor allowing for the simultaneous kinetic characterization of several multiplexed formats, such as 36 disparate antibodies targeting the same antigen, and facilitating detailed epitope binning and mapping studies. The kinetic rate constants determined by these methods correlated with those obtained on Biacore 2000 and the absolute parameter values obtained on the ProteOn's alginate-based GLC chip agreed closer with those from Biacore's flat C1 chip than Biacore's dextran-based CM4 chip. Pairwise epitope binning data from the ProteOn 36-ligand array format and those generated on an orthogonal array-based biosensor, the Octet QK384, gave similar results. In an epitope mapping study using biotinylated peptides, all three biosensor platforms were similar in their ability to identify antibodies that bound to linear epitopes. We apply alternative formats of the ProteOn array that enable a significantly higher number of assays to be conducted simultaneously than previously anticipated on this platform.
authors.affiliation: Rinat-Pfizer Inc., South San Francisco, CA 94080, USA. Yasmina.Abdiche@pfizer.com
authors.forename: Yasmina N
authors.initials: YN
authors.lastname: Abdiche
authors.forename: Kevin C
authors.initials: KC
authors.lastname: Lindquist
authors.forename: Alanna
authors.initials: A
authors.lastname: Pinkerton
authors.forename: Jaume
authors.initials: J
authors.lastname: Pons
authors.forename: Arvind
authors.initials: A
authors.lastname: Rajpal
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D055767:Immobilized Proteins / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D019151:Peptide Library; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D051381:Rats; D017186:Titrimetry
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D055767:Immobilized Proteins; D008024:Ligands; D019151:Peptide Library