Antigenic characterization of Brazilian isolates of Anaplasma marginale

Comparison between indirect enzyme-linked immunosorbent assays for Anaplasma marginale antibodies with recombinant major surface protein 5 and initial body antigens

“…Purification was certified by comparison in the sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) with noninfected bovine erythrocytes. Western blots with monoclonal antibodies (MoAbs) ANAF16C1 and ANA22B1, to MSP5 and MSP1a, respectively (Visser et al 1992, were also performed, as described elsewhere (Oliveira et al 2003).…”

“…The recombinant protein production was verified by SDS-PAGE, followed by Coomassie blue staining. The identity of rMSP5 was confirmed by Western blot with monoclonal antibody ANAF16C1, as described elsewhere (Oliveira et al 2003).…”

“…Recently, four new membrane proteins were identified: Ana29, Ana 32, Ana 37, and Ana 43 (Riding et al 2003). From these, the immunodominant 19-kDa MSP5 has potential use in serological tests (Ndung'u et al 1995, Knowles et al 1996, Reyna-Bello et al 1998, Torioni de Echaide 1998, as it is conserved between Anaplasma species and isolates, including Brazilian isolates of A. marginale (Visser et al 1992, Kano et al 2002, Oliveira et al 2003.…”

Progressos na imunização contra Anaplasma marginale

“…Uma única proteína MSP1a está ligada covalentemente, por pontes dissulfeto, a proteínas MSP1b (Vidotto et al 1994). A MSP1a é codificada por um único gene (msp1a), porém possui marcante variação de tamanho entre os diferentes isolados de A. marginale (46 a 105 kDa), como verificado em isolados brasileiros desta riquétsia (Kano et al 2002, Oliveira et al 2003 (Fig. 2).…”

“…Isso resulta da presença de um número variável de blocos repetitivos de 28 a 29 aminoácidos, ricos em serina, na região amino (N) terminal , Allred et al 1990). Essa região contém um epitopo sensível à neutralização conservado entre os isolados , Allred et al 1990), incluindo brasileiros (Kano et al 2002, Oliveira et al 2003) o qual é reconhecido pelos anticorpos monoclonais ANA22B1 e ANA15D2 , Allred et al 1990.…”