Journal
ACS SENSORS
Volume 1, Issue 5, Pages 614-622Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.6b00014
Keywords
nanopore; OmpG; sensing; biotin; antibody; streptavidin
Funding
- University of Massachusetts as part of the Chemistry-Biology Interface training grant [T32 GM08515]
- National Institutes of Health [R01GM115442]
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We have previously shown that a biotin ligand tethered to the rim of an OmpG nanopore can be used to detect biotin-binding proteins. Here, we investigate the effect of the length of the polyethylene glycol tether on the nanopore's sensitivity and selectivity. When the tether length was increased from 2 to 45 ethylene repeats, sensitivity decreased substantially for a neutral protein streptavidin and slightly for a positively charged protein (avidin). In addition, we found that two distinct avidin binding conformations were possible when using a long tether. These conformations were sensitive to the salt concentration and applied voltage. Finally, a longer tether resulted in reduced sensitivity due to slower association for a monoclonal antibiotin antibody. Our results highlight the importance of electrostatic, electroosmotic, and electrophoretic forces on nanopore binding kinetics and sensor readout.
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