2 research outputs found
The complementarity-determining region sequences in IgY antivenom hypervariable regions
The data presented in this article are related to the research article entitled ''Development of IgY antibodies against anti-snake toxins endowed with highly lethal neutralizing activity'' (da Rocha et al., 2017) [1]. Complementarity-determining region (CDR) sequences are variable antibody (Ab) sequences that respond with specificity, duration and strength to identify and bind to antigen (Ag) epitopes. B lymphocytes isolated from hens immunized with Bitis arietans (Ba) and anti-Crotalus durissus terrificus (Cdt) venoms and expressing high specificity, affinity and toxicity neutralizing antibody titers were used as DNA sources. The VLF1, CDR1, CDR2, VLR1 and CDR3 sequences were validated by BLASTp, and values corresponding to IgY VL and VH anti-Ba or anti-Cdt venoms were identified, registered [Gallus gallus IgY Fv Light chain (GU815099)/Gallus gallus IgY Fv Heavy chain (GU815098)] and used for molecular modeling of IgY scFv anti-Ba. The resulting CDR1, CDR2 and CDR3 sequences were combined to construct the three - dimensional structure of the Ab paratope. Keywords: PCR, Sequencing, Modeling of biomolecule
Identification of the apa protein secreted by Mycobacterium avium subsp. paratuberculosis as a novel fecal biomarker for Johne's disease in cattle
Paratuberculosis (PTB) or Johne's disease is a chronic intestinal infection of ruminants, caused by Mycobacterium avium subsp. paratuberculosis. The shedding of mycobacteria in the feces starts at the initial stages and increases with disease progression, suggesting that antigens secreted by mycobacteria could be excreted in the feces. Previously, we demonstrated that the alanine and proline-rich antigen (Apa), a secretory antigen of Map, could be detected in the intestine of cows with PTB using a monoclonal antibody. In this study, we verified whether this protein can be found in consistently detectable levels in the feces of cattle with PTB. Feces were obtained from cows with Johne's disease confirmed by laboratory tests, cows with suspected PTB based on seropositivity and from PTB-free control cows. Samples were immunoprecipitated using anti-Apa monoclonal antibody and analyzed by immunoblot. The Apa was detected as a 60/70 kDa doublet band in all samples obtained from animals with laboratory-confirmed disease and in a substantial proportion of seropositive asymptomatic animals, but not in the control samples. Additionally, the antigen was detected in the feces of animals with Johne's disease by ELISA. This study strongly suggests that Apa is a potential fecal biomarker of Johne's disease that could serve for immunodiagnosis.Fil: Da Silva De Souza, Giliane. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; BrasilFil: Rodriguez, Ana Barbara F.. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; BrasilFil: Romano, Maria Isabel. Instituto Nacional de TecnologÃa Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentina. Consejo Nacional de Investigaciones CientÃficas y Técnicas; ArgentinaFil: Ribeiro, Efranci Seliprandy. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; BrasilFil: Oelemann, Walter M. R.. Universidade Federal do Rio de Janeiro; BrasilFil: Da Rocha, David Gitirana. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; BrasilFil: Da Silva, Wilmar Dias. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Lasunskaia, Elena B.. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; Brasi