title: Cellular assay optimization: part I: the use of large-scale transiently transfected cryobanks and introduction of a c-Myc tag to design a standardized ELISA process.
abstract: This study investigated the use of large-scale transiently transfected cryopreserved cells for medium-throughput cellular screening. The data generated indicated that preprepared transiently transfected cryobanks can be used for cell-based assays and in fact can greatly enhance the consistency of data generated by cellular screens. In addition to this, a generic enzyme-linked immunosorbent assay method was designed that introduced a c-Myc tag to four different targets and allowed all four cell assays to be run using a standardized process. These process improvements yielded cost savings and greatly reduced the required resource, as well as reducing timelines for developing cellular assays.
authors.affiliation: Marie A. Macmillan was contracted to AstraZeneca via Morson International, Adamson House, Centenary Way, Salford, Manchester, UK.
authors.forename: Marie A
authors.initials: MA
authors.lastname: Macmillan
authors.forename: David I
authors.initials: DI
authors.lastname: Fisher
authors.forename: Karen
authors.initials: K
authors.lastname: Roberts
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Orme
mesh_terms: D018929:Cell Culture Techniques; D002461:Cell Line, Transformed; D015925:Cryopreservation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards*; D057809:HEK293 Cells; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics* / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014162:Transfection
chemical_list: D047428:Protein Kinase Inhibitors; D016271:Proto-Oncogene Proteins c-myc; D011494:Protein Kinases