Detection of pertussis toxin (PTX) activity is instrumental in the development and manufacturing of pertussis vaccines. Here, the researchers describe Interference in Gαi-mediated Signal Transduction (iGIST), an animal-free kinetic bioassay for the detection of PTX, by measuring its effect on inhibitory G protein-coupled receptor (GPCR) signalling. iGIST is based on HEK293 cells and a luminescent 3'5'-cyclic adenosine monophosphate (cAMP) probe. iGIST has a low sensitivity threshold in the pg/mL range of PTX, surpassing by 100-fold in a parallel analysis via the currently used in vitro end-point technique to detect PTX. iGIST also detects PTX in complex samples, i.e., a commercial PTX-toxoid-containing pertussis vaccine that was spiked with an active PTX. iGIST has an objective digital readout and is observer-independent, offering prospects for automation. iGIST emerges as a promising method to detect PTX activity in the development and manufacturing of pertussis vaccines.
iGIST— a kinetic bioassay for pertussis toxin based on its effect on inhibitory GPCR signaling
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