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Luciani M, Armillotta G, Manna L, et al. Selection of a monoclonal antibody by ELISA, immunoblotting and Quartz Crystal Microbalance technology for immunohistochemical detection of Mycoplasma mycoides subsp. mycoides. Vet Ital. 2016;doi: 10.12834/VetIt.435.2084.2.
Luciani, M., Armillotta, G., Manna, L., Ciarelli, A., Di Febo, T., Krasteva, I., Pini, A., Sacchini, F., & D'Angelo, A. R. (2016). Selection of a monoclonal antibody by ELISA, immunoblotting and Quartz Crystal Microbalance technology for immunohistochemical detection of Mycoplasma mycoides subsp. mycoides. Veterinaria italiana, . https://doi.org/10.12834/VetIt.435.2084.2
Luciani, Mirella, et al. "Selection of a monoclonal antibody by ELISA, immunoblotting and Quartz Crystal Microbalance technology for immunohistochemical detection of Mycoplasma mycoides subsp. mycoides." Veterinaria italiana vol. (2016). doi: https://doi.org/10.12834/VetIt.435.2084.2
Luciani M, Armillotta G, Manna L, Ciarelli A, Di Febo T, Krasteva I, Pini A, Sacchini F, D'Angelo AR. Selection of a monoclonal antibody by ELISA, immunoblotting and Quartz Crystal Microbalance technology for immunohistochemical detection of Mycoplasma mycoides subsp. mycoides. Vet Ital. 2016 Jul 15; doi: 10.12834/VetIt.435.2084.2. Epub 2016 Jul 15. PMID: 27723054.
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Vet Ital. 2016 Jul 15; doi: 10.12834/VetIt.435.2084.2. Epub 2016 Jul 15.

Selection of a monoclonal antibody by ELISA, immunoblotting and Quartz Crystal Microbalance technology for immunohistochemical detection of Mycoplasma mycoides subsp. mycoides.

Veterinaria italiana

Mirella Luciani, Gisella Armillotta, Laura Manna, Antonella Ciarelli, Tiziana Di Febo, Ivanka Krasteva, Attilio Pini, Flavio Sacchini, Anna Rita D'Angelo

Affiliations

  1. OIE Reference Laboratory for CBPP, Istituto Zoopro lattico Sperimentale dell'Abruzzo e del Molise 'G. Caporale', Campo Boario, 64100 Teramo, Italy.

PMID: 27723054 DOI: 10.12834/VetIt.435.2084.2

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.

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