Literature/202212301234 biggest challenge of single-molecule methods is non-specific binding
- Source: [@walt2013Optical Methods for Single Molecule Detection and Analysis]
- Tags: #single-molecule #non-specific-binding
The biggest problem of single-molecule detection methods that rely on labeling specific proteins is that there will always be a "background floor" of labels that bound to the wrong protein or to the substrate.
Non-specific binding (NSB) can be worked around by coating the substrates in such a way that there can be a washing step that limits the amount of background. However, it still poses the biggest limitation for single-molecule methods.
Pushing the detection limit down to single-molecules (see: why pushing sensitivity of assays) is very promising, but must be done carefully. The interpretation of the results must be done carefully.
This point is what makes the label-free methods such a relevant tool to characterize samples at low concentrations, focusing on the intrinsic properties of the analytes.
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