Nanobody-functionalized biological nanopores and means and methods related thereto

The invention relates to means and methods for analysis of analytes using nanopore-based sensors, for example, to methods, nanopore systems and devices for the stochastic detection of (label-free) analytes in complex samples, for example the specific detection of a protein biomarker in a bodily sample. Provided is a method for detecting the presence of at least one analyte in a sample using a nanopore system comprising a cis chamber comprising a first conductive liquid medium in liquid communication with a trans chamber comprising a second conductive liquid medium through a modified nanopore, comprising: (a) adding a sample to be analyzed for the presence of an analyte to the cis chamber; (b) optionally applying an electrical potential across the modified nanopore; (c) measuring ionic current passing through the modified nanopore, wherein said modified nanopore is a biological nanopore that is functionalized with a 5 to 50 kDa, preferably 10 to 40 kDa, recognition element (e.g., proteinaceous recognition element) R capable of specifically binding to the analyte.