title: Applicability of the Monocyte Activation Test (MAT) for hyperimmune sera in the routine of the quality control laboratory: Comparison with the Rabbit Pyrogen Test (RPT).
abstract: Pyrogen tests are safety assays performed during the routine quality control of injectable products required by regulatory agencies. Currently, there are three available testing possibilities: 1) the Rabbit Pyrogen Test (RPT); 2) the Bacterial Endotoxin Test (BET); and 3) test systems using human whole-blood or monocytes, termed Monocyte Activation Test (MAT). Although BET is often considered as a replacement for the animal test, it is unable to detect pyrogens other than endotoxin. MAT is based on the human fever reaction and thus, most closely reflects the human response. The aim of this study was to conduct a parallel comparison of the RPT and MAT for hyperimmune sera (HS) batches analyzed during the routine of a quality control laboratory. MAT was performed in the same 43 batches of HS previously tested using RPT. The results showed that MAT presented 100% sensitivity and approximately 85% specificity as compared to RPT, i.e., no false-negative results were obtained. Few suspicious samples, which were negative in the RPT after retesting, provided divergent positive results suggesting a lower limit of detection of MAT. MAT is thus able to detect contaminants in biological products such as HS batches.
authors.affiliation: Department of Pharmacology and Toxicology, National Institute of Quality Control in Health (INCQS), Oswaldo Cruz Foundation, Rio de Janeiro, Brazil; Centre for the Validation of Alternative Methods (BraCVAM), National Institute of Quality Control in Health (INCQS), Oswaldo Cruz Foundation, Rio de Janeiro, Brazil. Electronic address: cristiane.caldeira@incqs.fiocruz.br.
authors.forename: Cristiane Caldeira
authors.initials: CC
authors.lastname: da Silva
authors.affiliation: Department of Pharmacology and Toxicology, National Institute of Quality Control in Health (INCQS), Oswaldo Cruz Foundation, Rio de Janeiro, Brazil; Centre for the Validation of Alternative Methods (BraCVAM), National Institute of Quality Control in Health (INCQS), Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.
authors.forename: Octavio Augusto França
authors.initials: OA
authors.lastname: Presgrave
authors.affiliation: Johns Hopkins University, Bloomberg School of Public Health, Center for Alternatives to Animal Testing (CAAT), Baltimore, USA; University of Konstanz, CAAT-Europe, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hartung
authors.affiliation: Laboratory of Taxonomy, Biochemistry and Bioprospection of Fungi, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.
authors.forename: Aurea Maria Lage
authors.initials: AM
authors.lastname: de Moraes
authors.affiliation: National Institute of Quality Control in Health (INCQS), Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.
authors.forename: Isabella Fernandes
authors.initials: IF
authors.lastname: Delgado
mesh_terms: D000826:Animal Testing Alternatives; D000818:Animals; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology*; D053583:Interleukin-1beta / Q000276:immunology; D015850:Interleukin-6 / Q000276:immunology; D007753:Laboratories; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008297:Male; D009000:Monocytes / Q000187:drug effects* / Q000276:immunology; D011749:Pyrogens / Q000494:pharmacology*; D011786:Quality Control; D011817:Rabbits; D012680:Sensitivity and Specificity
chemical_list: D007106:Immune Sera; D053583:Interleukin-1beta; D015850:Interleukin-6; D008070:Lipopolysaccharides; D011749:Pyrogens
keywords: Endotoxin; Hyperimmune sera; Monocyte Activation Test; Rabbit Pyrogen Test