Botulinum toxin type B micromechanosensor.
Botulinum neurotoxin (BoNT) types A, B, E, and F are toxic to humans; early and rapid detection is essential for adequate medical treatment. Presently available tests for detection of BoNTs, although sensitive, require hours to days. We report a BoNT-B sensor whose properties allow detection of BoNT-B within minutes. The technique relies on the detection of an agarose bead detachment from the tip of a micromachined cantilever resulting from BoNT-B action on its substratum, the synaptic protein synaptobrevin 2, attached to the beads. The mechanical resonance frequency of the cantilever is monitored for the detection. To suspend the bead off the cantilever we use synaptobrevin's molecular interaction with another synaptic protein, syntaxin 1A, that was deposited onto the cantilever tip. Additionally, this bead detachment technique is general and can be used in any displacement reaction, such as in receptor-ligand pairs, where the introduction of one chemical leads to the displacement of another. The technique is of broad interest and will find uses outside toxicology.
Department of Physics, University of California, Riverside, CA 92521, USA.
W
W
Liu
Vedrana
V
Montana
Edwin R
ER
Chapman
U
U
Mohideen
Vladimir
V
Parpura
D000954:Antigens, Surface / Q000737:chemistry / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D015153:Blotting, Western; D001905:Botulinum Toxins / Q000737:chemistry* / Q000378:metabolism*; D019274:Botulinum Toxins, Type A; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007120:Immunochemistry; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D018625:Microscopy, Atomic Force / Q000295:instrumentation; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D050683:R-SNARE Proteins; D011994:Recombinant Proteins / Q000378:metabolism; D012680:Sensitivity and Specificity; D050827:Syntaxin 1; D013997:Time Factors
D000954:Antigens, Surface; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D050683:R-SNARE Proteins; D011994:Recombinant Proteins; D050827:Syntaxin 1; C096323:rimabotulinumtoxinB; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A