Fig. 2: Hydrodynamic force spectroscopy of the Biotin-Streptavidin and digoxigenin (DIG)-AntiDIG binding interaction.
From: Low cost and massively parallel force spectroscopy with fluid loading on a chip
A(i) Schematic of the setup used to examine the dissociation force of DIG and Anti-DIG binding interaction. The tether included a DIG end that enables binding to beads functionalized with Anti-DIG and a biotin end that enables binding of the tether to the streptavidin on the coverslip surface. (ii) Cumulative probability histogram of the recorded rupture forces under three different loading rates and (iii) the most probable rupture force (F*) with respect to applied loading rate (\(\dot{{{{{{\rm{F}}}}}}}\)) for DIG-AntiDIG binding interaction. N=3 most probable rupture forces were estimated based on random selection without replication of the recorded rupture force data under each loading rate. The estimated F* values are presented as mean values ±SD. B(i) Schematic of the setup used to execute hydrodynamic force spectroscopy on biotin-streptavidin binding interaction. The tether included two biotin ends that enable binding of the tether to beads functionalized with streptavidin and to the streptavidin on the coverslip surface. (ii) Cumulative probability histogram of the recorded rupture forces under three different loading rates and (iii) F* with respect to the applied \(\dot{{{{{{\rm{F}}}}}}}\) for biotin-streptavidin binding interaction. N = 3 most probable rupture forces were estimated based on random selection without replication of the recorded rupture force data under each loading rate. The estimated F* values are presented as mean values ±SD. For both binding interactions tested, the recorded rupture forces increased with increase in the applied \(\dot{{{{{{\rm{F}}}}}}}\) under semilogarithmic scale. The linear fit performed (black dash lines) indicated \(\triangle {{{{{\rm{X}}}}}}\) of 0.76 ± 0.05 nm and 0.95 ± 0.02 nm along with \({{{{{{\rm{k}}}}}}}_{{{{{{\rm{off}}}}}}}\) of 4.0 ± 1.4 × 10−4 s−1 and 4.7 ± 2.6 × 10−6 s−1 for DIG-AntiDIG and biotin-streptavidin binding interactions, respectively. Furthermore, the extracted energy landscape coefficients for biotin-streptavidin were used to project the most probable rupture force with respect to loading rate for a single biotin-streptavidin binding interaction (green dash line). Source data are provided as a Source Data file.
