TY  - JOUR
A1  - Pathe-Neuschäfer-Rube, Andrea
A1  - Neuschäfer-Rube, Frank
A1  - Haas, Gerald
A1  - Langoth-Fehringer, Nina
A1  - Püschel, Gerhard Paul
T1  - Cell-Based Reporter Release Assay to Determine the Potency of Proteolytic Bacterial Neurotoxins
JF  - Toxins
N2  - Despite the implementation of cell-based replacement methods, the mouse lethality assay is still frequently used to determine the activity of botulinum toxin (BoNT) for medical use. One explanation is that due to the use of neoepitope-specific antibodies to detect the cleaved BoNT substrate, the currently devised assays can detect only one specific serotype of the toxin. Recently, we developed a cell-based functional assay, in which BoNT activity is determined by inhibiting the release of a reporter enzyme that is liberated concomitantly with the neurotransmitter from neurosecretory vesicles. In theory, this assay should be suitable to detect the activity of any BoNT serotype. Consistent with this assumption, the current study shows that the stimulus-dependent release of a luciferase from a differentiated human neuroblastoma-based reporter cell line (SIMA-hPOMC1-26-GLuc cells) was inhibited by BoNT-A and-C. Furthermore, this was also inhibited by BoNT-B and tetanus toxin to a lesser extent and at higher concentrations. In order to provide support for the suitability of this technique in practical applications, a dose–response curve obtained with a pharmaceutical preparation of BoNT-A closely mirrored the activity determined in the mouse lethality assay. In summary, the newly established cell-based assay may represent a versatile and specific alternative to the mouse lethality assay and other currently established cell-based assays.
KW  - botulinum toxin
KW  - BoNT
KW  - tetanus toxin
KW  - RRR
KW  - replacement
Y1  - 2018
U6  - https://doi.org/10.3390/toxins10090360
SN  - 2072-6651
VL  - 10
IS  - 9
SP  - 1
EP  - 10
PB  - Molecular Diversity Preservation International (MDPI)
CY  - Basel
ER  -