title: Selection of a monoclonal antibody by ELISA, immunoblotting and Quartz Crystal Microbalance technology for immunohistochemical detection of Mycoplasma mycoides subsp. mycoides.
abstract: An immunohistochemical (IHC) technique was optimised using a monoclonal antibody (MAb) to detect Mycoplasma mycoides subsp. mycoides (Mmm), the agent of Contagious Bovine Pleuropneumonia (CBPP), in sections of lung tissue. A panel of MAbs was produced and screened for Mmm speci city and for cross-reactivity against other mycoplasmas belonging and not belonging to the Mycoplasma mycoides cluster, using in parallel indirect ELISA (i-ELISA) and Immunoblotting (IB). Based on i-ELISA and IB characterization data, 1 MAb (clone 3G10E7) was selected and its highest a nity vs Mmm was con rmed by the Quartz Crystal Microbalance (QCM) technology. Afterwards, IHC analyses were conducted to compare MAb 3G10E7 vs rabbit Mmm speci c hyperimmune serum using lung tissue sections of CBPP infected and CBPP negative animals. Results suggest that screening of MAbs using in parallel ELISA, IB, and QCM technology enables to select high a nity target speci c MAbs. Immunohistochemical results demonstrated that MAb 3G10E7 improved IHC performances, showing reduced background staining and no cross-reactivity against Mycoplasma bovis, which is responsible of pneumonia in cattle.
authors.affiliation: OIE Reference Laboratory for CBPP, Istituto Zoopro lattico Sperimentale dell'Abruzzo e del Molise 'G. Caporale', Campo Boario, 64100 Teramo, Italy.
authors.forename: Mirella
authors.initials: M
authors.lastname: Luciani
authors.forename: Gisella
authors.initials: G
authors.lastname: Armillotta
authors.forename: Laura
authors.initials: L
authors.lastname: Manna
authors.forename: Antonella
authors.initials: A
authors.lastname: Ciarelli
authors.forename: Tiziana
authors.initials: T
authors.lastname: Di Febo
authors.forename: Ivanka
authors.initials: I
authors.lastname: Krasteva
authors.forename: Attilio
authors.initials: A
authors.lastname: Pini
authors.forename: Flavio
authors.initials: F
authors.lastname: Sacchini
authors.forename: Anna Rita
authors.initials: AR
authors.lastname: D'Angelo