Seleção e caracterização de peptídeos recombinantesligantes a anticorpos monoclonais reativos a proteínas de Anaplasma marginale
Bovine anaplasmosis is caused by Anaplasma marginale and A.centrale. The most pathogenic and important species for cattle production is A.marginale, and is widely distributed in tropical, subtropical and temperateregions of the world. A. marginale is an intra-erythrocyte rickettsia of susceptibleruminants, biological and mechanically transmitted by ticks and hematophagousinsects. The tick Rhipicephalus (Boophilus) microplus is the main vector of A.marginale in Brazil. The congenital form of transmission in cattle may occur,causing the neonatal anaplasmosis. The outer membrane of A. marginaleincludes six well characterized major surface proteins, MSP1a, MSP1b, MSP2,MSP3, MSP4 and MSP5, which play important role in the development of theimmune response of infected animals. In this study, we have used the PhageDisplay technology to identify specific peptides that were immunoreactive tomonoclonal antibodies anti-A. marginale proteins. Peptide selection wasperformed using a subtractive selection of a peptide library with 12 randomamino acids, Ph.D.-12, expressed on the surface of the M13 filamentous phageconcurrently against the anti-MSP1a and anti-MSP2. After four rounds ofselection and validation by ELISA, the selected peptides have recognized onlythe anti-MSP1. Analysis of bioinformatics identified 45 peptides, which showedthe protein consensus sequence STxS that was represented in 78% of selectedphages. Due to the multiple motif repeats found in MSP1 protein, the STSSxLmotif may become an important biological target, with potential use in diagnostictests and vaccine for the control of Anaplasma marginale.
Advisor:Luiz Ricardo Goulart Filho; Malcon Antonio Manfredi Brandeburgo; Carlos Henrique Marchiori; Guilherme Rocha Lino de Souza
School:Universidade Federal de Uberlândia
Source Type:Master's Thesis
Date of Publication:06/30/2008