title: Multiplex immunoassay for in vitro characterization of acellular pertussis antigens in combination vaccines.
abstract: Vaccines characterization is required to ensure physical, chemical, and biological integrity of antigens and adjuvants. Current analytical methods mostly require complete antigen desorption from aluminum-based adjuvants and are not always suitable to distinguish individual antigens in multivalent formulations. Here, Luminex technology is proposed to improve the analytics of vaccine characterization. As proof of concept, TdaP (tetanus, diphtheria and acellular pertussis) combination, adjuvanted with aluminum hydroxide, was chosen as model formulation to quantify and determine the level of adsorption of acellular pertussis (aP) antigens onto adjuvant surface at the same time. The assay used specific antibodies bound to magnetic microspheres presenting unique digital signatures for each pertussis antigen, allowing the simultaneous recognition of respective antigens in the whole vaccine, avoiding laborious procedures for adjuvant separation. Accurate and reproducible quantification of aP antigens in TdaP vaccine has been achieved in the range 0.78-50 ng/mL, providing simultaneously information on antigen identity, quantity, and degree of adsorption to aluminum hydroxide. The current study could further be considered as a model to set up in vitro potency assays thus supporting the replacement of animal tests accordingly to the 3Rs concept.
authors.affiliation: Novartis Vaccines and Diagonstics SrL (a GSK Company), Via Fiorentina 1, 53100 Siena, Italy; Università degli Studi di Padova, 35129 Padova, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Agnolon
authors.affiliation: Novartis Vaccines and Diagonstics SrL (a GSK Company), Via Fiorentina 1, 53100 Siena, Italy; Università di Siena, 53100 Siena, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Bruno
authors.affiliation: Novartis Vaccines and Diagonstics SrL (a GSK Company), Via Fiorentina 1, 53100 Siena, Italy.
authors.forename: Bruno
authors.initials: B
authors.lastname: Galletti
authors.affiliation: Novartis Vaccines and Diagonstics SrL (a GSK Company), Via Fiorentina 1, 53100 Siena, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Mori
authors.affiliation: GlaxoSmithKline (former Novartis Vaccines), 350 Massachusetts Ave, Cambridge, 02139 MA, USA.
authors.forename: Mildred
authors.initials: M
authors.lastname: Ugozzoli
authors.affiliation: Novartis Vaccines and Diagonstics SrL (a GSK Company), Via Fiorentina 1, 53100 Siena, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Pergola
authors.affiliation: GlaxoSmithKline (former Novartis Vaccines), 350 Massachusetts Ave, Cambridge, 02139 MA, USA.
authors.forename: Derek T
authors.initials: DT
authors.lastname: O'Hagan
authors.affiliation: Novartis Vaccines and Diagonstics SrL (a GSK Company), Via Fiorentina 1, 53100 Siena, Italy. Electronic address: barbara.c.baudner@gsk.com.
authors.forename: Barbara C
authors.initials: BC
authors.lastname: Baudner
mesh_terms: D018829:Adhesins, Bacterial / Q000737:chemistry; D000276:Adjuvants, Immunologic / Q000737:chemistry*; D000942:Antigens, Bacterial / Q000737:chemistry*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D008863:Microspheres; D037342:Pertussis Toxin / Q000737:chemistry; D010567:Pertussis Vaccine / Q000737:chemistry*; D017778:Vaccines, Combined / Q000737:chemistry; D010566:Virulence Factors, Bordetella / Q000737:chemistry
chemical_list: D018829:Adhesins, Bacterial; D000276:Adjuvants, Immunologic; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D010567:Pertussis Vaccine; D017778:Vaccines, Combined; D010566:Virulence Factors, Bordetella; C065273:filamentous hemagglutinin adhesin, Bordetella pertussis; C068929:pertactin; D037342:Pertussis Toxin
keywords: Acellular pertussis; Aluminum hydroxide; Combination vaccine; Luminex technology; Multiplex