title: Development and validation of an enzyme linked immunosorbent assay for palivizumab serum determination.
abstract: Palivizumab (Synagis) is a humanized monoclonal antibody (IgG1K) composed of 95 percent human and 5 percent murine sequences. It is directed to an epitope in the A antigenic site of the F protein of respiratory syncytial virus (RSV). Palivizumab is used for prevention of serious lower respiratory tract disease caused by RSV in pediatric patients who are at increased risk of severe disease and is administered intramuscularly (IM) for a total of 5 monthly doses. Herein, we report on the development and validation of a very sensitive enzyme-linked immunosorbent assay (ELISA) to measure serum concentrations of palivizumab by a rabbit polyclonal antibody specifically produced against the murine sequence. The method was developed and validated according to the guidelines "Guidance for Industry" (1998) and has proved suitable for the determination of palivizumab serum levels in the target infant population. The ELISA assay was successfully applied to test the serum samples in an infant population who received palivizumab intramuscularly; thus, the assay could be used to determine serum levels in palivizumab-treated infants to optimize dosing and scheduling and to study the relationship between dose and clinical response.
authors.affiliation: Clinical Pharmacokinetics and Experimental Unit, Foundation IRCCS Policlinico San Matteo, Pavia, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Montagna
authors.forename: L
authors.initials: L
authors.lastname: Visai
authors.forename: A
authors.initials: A
authors.lastname: Di Comite
authors.forename: V
authors.initials: V
authors.lastname: Iommiello
authors.forename: M A
authors.initials: MA
authors.lastname: Avanzini
authors.forename: N
authors.initials: N
authors.lastname: Bloise
authors.forename: M
authors.initials: M
authors.lastname: Stronati
authors.forename: M
authors.initials: M
authors.lastname: Ragazzi
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage / Q000097:blood*; D000998:Antiviral Agents / Q000008:administration & dosage / Q000097:blood*; D002138:Calibration; D016903:Drug Monitoring / Q000379:methods* / Q000592:standards; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D006801:Humans; D007223:Infant; D007273:Injections, Intramuscular; D057230:Limit of Detection; D016014:Linear Models; D015588:Observer Variation; D000069455:Palivizumab; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D000998:Antiviral Agents; D000069455:Palivizumab