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Highly sensitive protein detection by combination of atomic force microscopy fishing with charge generation and mass spectrometry analysis
Author(s) -
Ivanov Yuri D.,
Pleshakova Tatyana,
Malsagova Krystina,
Kozlov Andrey,
Kaysheva Anna,
Kopylov Arthur,
Izotov Alexander,
Andreeva Elena,
Kanashenko Sergey,
Usanov Sergey,
Archakov Alexander
Publication year - 2014
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.13011
Subject(s) - pyrolytic carbon , electric field , mass spectrometry , analytical chemistry (journal) , voltage , atomic force microscopy , chemistry , materials science , nanotechnology , chromatography , physics , organic chemistry , quantum mechanics , pyrolysis
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 b 5 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.