Highly sensitive protein detection by biospecific AFM-based fishing with pulsed electrical stimulation.
Tatyana O Pleshakova, Kristina A Malsagova, Anna L Kaysheva, Arthur T Kopylov, Vadim Yu Tatur, Vadim S Ziborov, Sergey L Kanashenko, Rafael A Galiullin, Yuri D Ivanov
Author Information
Tatyana O Pleshakova: Institute of Biomedical Chemistry Moscow Russia.
Kristina A Malsagova: Institute of Biomedical Chemistry Moscow Russia.
Anna L Kaysheva: Institute of Biomedical Chemistry Moscow Russia.
Arthur T Kopylov: Institute of Biomedical Chemistry Moscow Russia.
Vadim Yu Tatur: Foundation of Perspective Technologies and Novations Moscow Russia.
Vadim S Ziborov: Joint Institute for High Temperatures RAS Moscow Russia.
Sergey L Kanashenko: Institute of Biomedical Chemistry Moscow Russia.
Rafael A Galiullin: Institute of Biomedical Chemistry Moscow Russia.
Yuri D Ivanov: Institute of Biomedical Chemistry Moscow Russia.
We report here the highly sensitive detection of protein in solution at concentrations from 10 to 10 m using the combination of atomic force microscopy (AFM) and mass spectrometry. Biospecific detection of biotinylated bovine serum albumin was carried out by fishing out the protein onto the surface of AFM chips with immobilized avidin, which determined the specificity of the analysis. Electrical stimulation was applied to enhance the fishing efficiency. A high sensitivity of detection was achieved by application of nanosecond electric pulses to highly oriented pyrolytic graphite placed under the AFM chip. A peristaltic pump-based flow system, which is widely used in routine bioanalytical assays, was employed throughout the analysis. These results hold promise for the development of highly sensitive protein detection methods using nanosensor devices.
