Hydrophobin-coated plates as matrix-assisted laser desorption/ionization sample support for peptide/protein analysis |
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Authors: | Sara Longobardi Alfredo Maria Gravagnuolo Ilaria Rea Luca De Stefano Gennaro Marino Paola Giardina |
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Institution: | 1. Department of Chemical Sciences, University of Naples “Federico II”, 80126 Naples, Italy;2. Unit of Naples–Institute for Microelectronics and Microsystems, National Council of Research, 80131 Naples, Italy |
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Abstract: | Fungal hydrophobins are amphipathic self-assembling proteins. Vmh2 hydrophobin, prepared from mycelial cultures of the basidiomycete fungus Pleurotus ostreatus, spontaneously forms a stable and homogeneous layer on solid surfaces and is able to strongly absorb proteins even in their active forms. In this work, we have exploited the Vmh2 self-assembled layer as a novel coating of a matrix-assisted laser desorption/ionization (MALDI) steel sample-loading plate. Mixtures of standard proteins, as well as tryptic peptides, in the nanomolar–femtomolar range were analyzed in the presence of salts and denaturants. As evidence on a real complex sample, crude human serum was also analyzed and spectra over a wide mass range were acquired. A comparison of this novel coating method with both standard desalting techniques and recently reported on-plate desalting methods was also performed. The results demonstrate that Vmh2 coating of MALDI plates allows for a very simple and effective desalting method suitable for development of lab-on-a-plate platforms focused on proteomic applications. |
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Keywords: | Thin films Fungal proteins Self-assembly Lab-on-a-plate |
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