title: Thin-layer matrix sublimation with vapor-sorption induced co-crystallization for sensitive and reproducible SAMDI-TOF MS analysis of protein biosensors.
abstract: Coupling immunoassays on self-assembled monolayers (SAMs) to matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) provides improved assay selectivity compared with traditional photometric detection techniques. We show that thin-layer-transfer (TLT) of α-cyano-4-hydroxycinnaminic acid (CHCA) MALDI matrix via vacuum sublimation followed by organic solvent-based vapor-sorption induced co-crystallization (VIC) results in unique matrix/analyte co-crystallization tendencies that optimizes assay reproducibility and sensitivity. Unique matrix crystal morphologies resulted from VIC solvent vapors, indicating nucleation and crystal growth characteristics depend upon VIC parameters. We observed that CHCA microcrystals generated by methanol VIC resulted in >10× better sensitivity, increased analyte charging, and improved precision compared with dried droplet measurements. The uniformity of matrix/analyte co-crystallization across planar immunoassays directed at intact proteins yielded low spectral variation for single shot replicates (18.5 % relative standard deviation, RSD) and signal averaged spectra (<10 % RSD). We envision that TLT and VIC for MALDI-TOF will enable high-throughput, reproducible array-based immunoassays for protein molecular diagnostic assays in diverse biochemical and clinical applications.
authors: Department of Pathology, University of Texas Southwestern Medical Center, Dallas, 75390, USA.
authors: Michael J
authors: MJ
authors: Roth
authors: Jaekuk
authors: J
authors: Kim
authors: Erica M
authors: EM
authors: Maresh
authors: Daniel A
authors: DA
authors: Plymire
authors: John R
authors: JR
authors: Corbett
authors: Junmei
authors: J
authors: Zhang
authors: Steven M
authors: SM
authors: Patrie
mesh_terms: D000327:Adsorption; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002417:Cattle; D003460:Crystallization; D006736:Horses; D006801:Humans; D055767:Immobilized Proteins; D007118:Immunoassay / Q000379:methods; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D055767:Immobilized Proteins; D011506:Proteins