INVESTIGADORES
GRIPPO vanina
congresos y reuniones científicas
Título:
OBTAINING AND CHARACTERIZING OF SINGLE DOMAIN ANTIBODIES DERIVED FROM CAMELIDS (NANOANTIBODIES OR VHH) AGAINST FOOT-AND-MOUTH DISEASE VIRUS FOR DIAGNOSTIC APPLICATION
Autor/es:
JOAQUIN BOZZO; FLORENCIA MARCHESE; NORA MATTION; VANINA GRIPPO
Lugar:
CABA
Reunión:
Congreso; JOINT MEETING OF BIOSCIENCE SOCIETIES; 2017
Institución organizadora:
LXII ANNUAL MEETING OF ARGENTINE SOCIETY OF CLINICAL INVESTIGATION (SAIC)
Resumen:
Foot-and-Mouth Disease (FMD) is a viral disease of livestock with serious economic repercussions. Development of innovative diagnostic methods for the FMD control is of concern due to the susceptibility of species of economic interest. Camelids present heavy chain only antibodies (IgG 2 and 3), so the variable domain consists of a single domain (VHH or Nanoantibody). These antibodies represent a novel tool with unique characteristics conferring great potential in the development of new more sensitive diagnostic methods. For this reason, the present work aimed to obtain and preliminary characterize VHH specific against FMDV for its application in the diagnostic innovation. Two llamas (Lama glama) were immunized with the strains O1/Campos, A24/Cruzeiro (A24), A/Arg/2001 (A2001) and C3/Indaial (C3). Humoral immune response (total IgG and subtype titers) was assessed throughout the immunization schedule obtaining high levels of heavy chain-only antibodies against the 4 serotypes by ELISA. Serum reactivity against viral proteins was confirmed in Western Blot. Two Nanoantibody libraries were constructed starting from llama peripheral blood and VHH against O1C strain were selected by Phage Display. Even more, 119 out of 182 selected clones recognized the 4 serotypes in ELISA. From these clones, 17 which showed very high reactivity in ELISA were selected for diversity analysis by enzymatic digestion. As a result, it was shown that 16 out of 17 clones presented different fingerprinting pattern. In conclusion, two VHH libraries were constructed from immunized llama with the O1C, A24, A2001 and C3 strains. Both llamas had a high immune response and their sera were able to recognize the 4 FMDV serotypes. Furthermore, the two llamas had high levels of heavy chain only antibodies, mainly IgG3 subtype. At least 16 different cross-reactive Nanoantibodies were obtained against the 4 strains. Thus, these antibodies represent a potential tool for diagnostic innovation of FMDV.