title: Highly sensitive protein detection by combination of atomic force microscopy fishing with charge generation and mass spectrometry analysis.
abstract: An approach combining atomic force microscopy (AFM) fishing and mass spectrometry (MS) analysis to detect proteins at ultra-low concentrations is proposed. Fishing out protein molecules onto a highly oriented pyrolytic graphite surface coated with polytetrafluoroethylene film was carried out with and without application of an external electric field. After that they were visualized by AFM and identified by MS. It was found that injection of solution leads to charge generation in the solution, and an electric potential within the measuring cell is induced. It was demonstrated that without an external electric field in the rapid injection input of diluted protein solution the fishing is efficient, as opposed to slow fluid input. The high sensitivity of this method was demonstrated by detection of human serum albumin and human cytochrome b5 in 10(-17) -10(-18) m water solutions. It was shown that an external negative voltage applied to highly oriented pyrolytic graphite hinders the protein fishing. The efficiency of fishing with an external positive voltage was similar to that obtained without applying any voltage.
authors.affiliation: Institute of Biomedical Chemistry RAMS, Moscow, Russia.
authors.forename: Yuri D
authors.initials: YD
authors.lastname: Ivanov
authors.forename: Tatyana
authors.initials: T
authors.lastname: Pleshakova
authors.forename: Krystina
authors.initials: K
authors.lastname: Malsagova
authors.forename: Andrey
authors.initials: A
authors.lastname: Kozlov
authors.forename: Anna
authors.initials: A
authors.lastname: Kaysheva
authors.forename: Arthur
authors.initials: A
authors.lastname: Kopylov
authors.forename: Alexander
authors.initials: A
authors.lastname: Izotov
authors.forename: Elena
authors.initials: E
authors.lastname: Andreeva
authors.forename: Sergey
authors.initials: S
authors.lastname: Kanashenko
authors.forename: Sergey
authors.initials: S
authors.lastname: Usanov
authors.forename: Alexander
authors.initials: A
authors.lastname: Archakov
mesh_terms: D015786:Cytochromes b5 / Q000032:analysis*; D006801:Humans; D018625:Microscopy, Atomic Force / Q000295:instrumentation* / Q000379:methods*; D010446:Peptide Fragments / Q000032:analysis*; D012709:Serum Albumin / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D010446:Peptide Fragments; D012709:Serum Albumin; D015786:Cytochromes b5
keywords: AFM; MS; protein fishing; ultra-low protein concentration