title: Surface preparation strategies for improved parallelization and reproducible MALDI-TOF MS ligand binding assays.
abstract: Immunoassays are employed in academia and the healthcare and biotech industries for high-throughput, quantitative screens of biomolecules. We have developed monolayer-based immunoassays for MALDI-TOF MS. To improve parallelization, we adapted the workflow to photolithography-generated arrays. Our work shows Parylene-C coatings provide excellent "solvent pinning" for reagents and biofluids, enabling sensitive MS detection of immobilized components. With a unique MALDI-matrix crystallization technique we show routine interassay RSD <10% at picomolar concentrations and highlight platform compatibility for relative and label-free quantitation applications. Parylene-arrays provide high sample densities and promise screening throughputs exceeding 1000 samples/h with modern liquid-handlers and MALDI-TOF systems.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Roth
authors.forename: Erica M
authors.initials: EM
authors.lastname: Maresh
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Plymire
authors.forename: Junmei
authors.initials: J
authors.lastname: Zhang
authors.forename: John R
authors.initials: JR
authors.lastname: Corbett
authors.forename: Roger
authors.initials: R
authors.lastname: Robbins
authors.forename: Steven M
authors.initials: SM
authors.lastname: Patrie
mesh_terms: D006454:Hemoglobins / Q000737:chemistry; D007118:Immunoassay; D008024:Ligands; D011108:Polymers / Q000737:chemistry; D021122:Protein Subunits / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods* / Q000592:standards; D014992:Xylenes / Q000737:chemistry
chemical_list: D006454:Hemoglobins; D008024:Ligands; D011108:Polymers; D021122:Protein Subunits; D014992:Xylenes; C011055:parylene