|Common screening technologies for infectious agent detection in biopharmaceutical drug production employ agar or broth-culture techniques for bacteria including mycoplasma, and cell infectivity assays for detection of virus. In the case of virus detection, susceptible mammalian cell lines known to propagate specific viral species are monitored by visual readout of cytopathic effects. Cytopathic effects have a variety of phenotypic displays; cell death, onset of abnormal cellular morphology and formation of large multinucleated cells are common effects signaling a contaminating species. Such agar/broth culture assays and cell infectivity assays have been shown in certain instances to be less sensitive than newer technologies. For example, although detection of mycoplasma was achieved in test samples using nucleic acid-based techniques, the same mycoplasma was not detected in standard broth-culture assays or even in hybrid culture-qPCR assays. |
Thus sensitivity and broad pathogen detection are prerequisites to biotherapeutic drug safety. Technology which allows an unequivocal determination of any and every potential pathogen present in a production sample is an ultimate goal. Studies on new technologies being explored through collaboration between Amgen and companies engaged in drug safety technologies are ongoing. For example, technology where PCR amplification is coupled to amplicon analysis by mass spectrometry may be a powerful tool for rapid detection of potential pathogens whether viral, bacterial, fungal or protozoa. Studies evaluating this technology will be presented.