“…However, apart from the mouse bioassay [39] and one sandwich ELISA [40], none of the other methods had been evaluated in a PT or method performance study until recently. A BoNT PT organized by the EQuATox consortium including 23 laboratories allowed the comparison of various methods and their performance [41]. In this exercise, high purity BoNT/A-F materials were generated and [21,[24][25][26] In vitro cytotoxicity tests Measurement of cell death in vitro (endpoint and real-time assays) [21,27] BoNT/A, B and E Immunological assays Sandwich ELISA: Detection of antigen by a combination of mAb and/or pAb, i) four combinations of two pAb each from goat and/or horse targeting BoNT/A, B, E, or F; detection of holotoxin [40] ii) three combinations of two mAb targeting BoNT/A (mAb TA18 + mAb TA13), BoNT/B (mAb TB6 + mAb TB17), or BoNT/E (mAb TE81 + mAb TE49); detection of HC-domain [42,107] iii) three combinations of mAb and/or pAb targeting BoNT/A (mAb A1688 + mAb HcA78), BoNT/B (mAb B279 + horse trivalent pAb), or BoNT/E (rabbit pAb KE97 + mAb E136); detection of holotoxin [41,42,106] Lateral Flow assay: Sandwich ELISA on a paper strip i) three combinations of two mAb targeting A (mAb TA12 + mAb TA9), B (mAb TB6 + mAb TB17), or E (mAb TE81 + mAb TE49); detection of HC-domain [41,42] (continued on next page) Chromatographic methods…”