The alpha-hemolysin nanopore transduction detector - single-molecule binding studies and immunological screening of antibodies and aptamers.
BACKGROUND
Nanopore detection is based on observations of the ionic current threading a single, highly stable, nanometer-scale channel. The dimensions are such that small biomolecules and biopolymers (like DNA and peptides) can translocate or be captured in the channel. The identities of translocating or captured molecules can often be discerned, one from another, based on their channel blockade "signatures". There is a self-limiting aspect to a translocation-based detection mechanism: as the channel fits tighter around the translocating molecule the dynamic range of the ionic current signal is reduced. In this study, a lengthy, highly structure, high dynamic-range, molecular capture is sought as a key component of a transduction-based nanopore detection platform.


RESULTS
A specialized role, or device augmentation, involving bifunctional molecules has been explored. The bifunctional molecule has one function to enter and blockade the channel in an information-rich self-modulating manner, while the other function is for binding (usually), located on a non-channel-captured portion of the molecule. Part of the bifunctional molecule is, thus, external to the channel and is free to bind or rigidly link to a larger molecule of interest. What results is an event transduction detector: molecular events are directly transduced into discernible changes in the stationary statistics of the bifunctional molecule's channel blockade. Several results are presented of nanopore-based event-transduction detection.


CONCLUSIONS
It may be possible to directly track the bound versus unbound state of a huge variety of molecules using nanopore transduction detection.
Research Institute for Children, Children's Hospital, 200 Henry Clay Ave, New Orleans, LA 70118, USA. winters@cs.uno.edu
Stephen
S
Winters-Hilt
D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D052157:Aptamers, Nucleotide / Q000032:analysis* / Q000276:immunology; D001427:Bacterial Toxins / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006460:Hemolysin Proteins / Q000737:chemistry*; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods; D016062:Porosity; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014159:Transducers
D000906:Antibodies; D052157:Aptamers, Nucleotide; D001427:Bacterial Toxins; D006460:Hemolysin Proteins; C021091:staphylococcal alpha-toxin