Abstract:
:By combining optical tweezers-assisted dynamic force spectroscopy experiments with fluorescence activated cell sorting (FACS), we demonstrate a new approach to reducing the data variance in measuring receptor-ligand interactions on a single molecule level by ensuring similar coating densities. Therefore, the carboxyfluorescein-labelled monophosphorylated peptide tau226-240[pThr231] is anchored on melamine resin beads and these beads are sorted by FACS to achieve a homogeneous surface coverage. To quantify the impact of the fluorescence dye on the bond parameters between the phosphorylated peptide and the corresponding phosphorylation specific anti-human tau monoclonal antibody HPT-104, we perform dynamic force spectroscopy and compare the results to data using unsorted beads covered with the non-fluorescence peptide analogue. Finally, we demonstrate that the data variance of the relative binding frequency is significantly decreased by a factor of 3.4 using pre-sorted colloids with a homogeneous ligand coating compared to using unsorted colloids.
journal_name
Phys Bioljournal_title
Physical biologyauthors
Stangner T,Singer D,Wagner C,Gutsche C,Ueberschär O,Hoffmann R,Kremer Fdoi
10.1088/1478-3975/10/4/046004subject
Has Abstractpub_date
2013-08-01 00:00:00pages
046004issue
4eissn
1478-3967issn
1478-3975journal_volume
10pub_type
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