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Highly sensitive protein detection by combination of atomic force microscopy fishing with charge generation and mass spectrometry analysis
- Source :
- The FEBS journal. 281(20)
- Publication Year :
- 2013
-
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.
- Subjects :
- Polytetrafluoroethylene
Atomic force microscopy
Analytical chemistry
Cell Biology
Human serum albumin
Mass spectrometry
Microscopy, Atomic Force
Biochemistry
Peptide Fragments
chemistry.chemical_compound
Cytochromes b5
chemistry
Highly oriented pyrolytic graphite
Electric field
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
medicine
Humans
Electric potential
Molecular Biology
Serum Albumin
Voltage
medicine.drug
Subjects
Details
- ISSN :
- 17424658
- Volume :
- 281
- Issue :
- 20
- Database :
- OpenAIRE
- Journal :
- The FEBS journal
- Accession number :
- edsair.doi.dedup.....d2b518dad8c58935255f420b44ef12d3