title: A new surface plasmon resonance-based immunoassay for rapid, reproducible and sensitive quantification of pentraxin-3 in human plasma.
abstract: A new immunoassay based on surface plasmon resonance (SPR) for the rapid, reproducible and sensitive determination of pentraxin-3 (PTX3) levels in human plasma has been developed and characterized. The method involves a 3-min flow of plasma over a sensor chip pre-coated with a monoclonal anti-PTX3 antibody (MNB4), followed by a 3-min flow of a polyclonal anti-PTX3 antibody (pAb), required for specific recognition of captured PTX3. The SPR signal generated with this secondary antibody linearly correlates with the plasma PTX3 concentration, in the range of 5-1500 ng/mL, with a lowest limit of detection of 5 ng/mL. The PTX3 concentrations determined with the SPR-based immunoassay in the plasma of 21 patients with sepsis, ranging 15-1600 ng/mL, were superimposable to those found in a classic ELISA immunoassay. Since the PTX3 concentration in the plasma of healthy subjects is <2 ng/mL, but markedly rises in certain medical conditions, the method is useful to quantify pathological levels of this important biomarker, with important diagnostic applications. In comparison with the classic ELISA, the SPR-based approach is much faster (30 min versus 4-5 h) and could be exploited for the development of new cost-effective SPR devices for point-of-care diagnosis.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Milan 20156, Italy. mara.canovi@gmail.com.
authors.forename: Mara
authors.initials: M
authors.lastname: Canovi
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Milan 20156, Italy. jacopo.lucchetti@marionegri.it.
authors.forename: Jacopo
authors.initials: J
authors.lastname: Lucchetti
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Milan 20156, Italy. matteo.stravalaci@marionegri.it.
authors.forename: Matteo
authors.initials: M
authors.lastname: Stravalaci
authors.affiliation: Humanitas Clinical and Research Hospital, Rozzano, Milan 20089, Italy. Sonia.Valentino@humanitasresearch.it.
authors.forename: Sonia
authors.initials: S
authors.lastname: Valentino
authors.affiliation: Humanitas Clinical and Research Hospital, Rozzano, Milan 20089, Italy. Barbara.Bottazzi@humanitasresearch.it.
authors.forename: Barbara
authors.initials: B
authors.lastname: Bottazzi
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Milan 20156, Italy. matteo.stravalaci@marionegri.it.
authors.forename: Mario
authors.initials: M
authors.lastname: Salmona
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Milan 20156, Italy. antonio.bastone@marionegri.it.
authors.forename: Antonio
authors.initials: A
authors.lastname: Bastone
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Milan 20156, Italy. marco.gobbi@marionegri.it.
authors.forename: Marco
authors.initials: M
authors.lastname: Gobbi
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D001774:Blood Chemical Analysis / Q000295:instrumentation*; D002097:C-Reactive Protein / Q000032:analysis*; D003199:Computer Systems; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D000683:Serum Amyloid P-Component / Q000032:analysis*; D020349:Surface Plasmon Resonance / Q000295:instrumentation*; D000099153:Pentraxins
chemical_list: D002097:C-Reactive Protein; D000683:Serum Amyloid P-Component; D000099153:Pentraxins