title: Fluorinated liquid-enabled protein handling and surfactant-aided crystallization for fully in situ digital microfluidic MALDI-MS analysis.
abstract: A droplet (digital) microfluidic device has been developed that enables complete protein sample preparation for MALDI-MS analysis. Protein solution dispensing, disulfide bond reduction and alkylation, tryptic digestion, sample crystallization, and mass spectrometric analysis are all performed on a single device without the need for any ex situ sample purification. Fluorinated solvents are used as an alternative to surfactants to facilitate droplet movement and limit protein adsorption onto the device surface. The fluorinated solvent is removed by evaporation and so does not interfere with the MALDI-MS analysis. Adding a small amount of perfluorooctanoic acid to the MALDI matrix solution improves the yield, quality and consistency of the protein-matrix co-crystals, reducing the need for extensive 'sweet spot' searching and improving the spectral signal-to-noise ratio. These innovations are demonstrated in the complete processing and MALDI-MS analysis of lysozyme and cytochrome c. Because all of the sample processing steps and analysis can be performed on a single digital microfluidic device without the need for ex situ sample handling, higher throughput can be obtained in proteomics applications. More generally, the results presented here suggest that fluorinated liquids could also be used to minimize protein adsorption and improve crystallization in other types of lab-on-a-chip devices and applications.
authors.affiliation: Biomedical Engineering Interdepartmental PhD Program, University of California, Los Angeles, CA 90095-1600, USA.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Aijian
authors.forename: Debalina
authors.initials: D
authors.lastname: Chatterjee
authors.forename: Robin L
authors.initials: RL
authors.lastname: Garrell
mesh_terms: D002210:Caprylates / Q000737:chemistry; D003460:Crystallization; D045304:Cytochromes c / Q000737:chemistry* / Q000378:metabolism; D005466:Fluorocarbons / Q000737:chemistry; D054879:Halogenation; D046210:Microfluidic Analytical Techniques; D009113:Muramidase / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000032:analysis; D059629:Signal-To-Noise Ratio; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013501:Surface-Active Agents / Q000737:chemistry*; D014357:Trypsin / Q000378:metabolism
chemical_list: D002210:Caprylates; D005466:Fluorocarbons; D010455:Peptides; D013501:Surface-Active Agents; D045304:Cytochromes c; C023036:perfluorooctanoic acid; D009113:Muramidase; D014357:Trypsin