Artigo de Periódico
Three-phase partitioning as an efficient method for extraction/concentration of immunoreactive excreted–secreted proteins of Corynebacterium pseudotuberculosis
Fecha
2004Registro en:
1046-5928
v. 34, n. 2
Autor
Paule, B. J. A.
Nascimento, Roberto José Meyer
Costa, L. F. Moura
Bahia, Robson Cerqueira
Carminati, R.
Regis, Lia Fernandes
Paule, B. J. A.
Nascimento, Roberto José Meyer
Costa, L. F. Moura
Bahia, Robson Cerqueira
Carminati, R.
Regis, Lia Fernandes
Institución
Resumen
The three-phase partitioning (TPP) technique was used upstream to isolate/concentrate secreted proteins from Corynebacterium
pseudotuberculosis cultured in a complex liquid medium. Several parameters of the TPP technique (15, 30, or 60% ammonium sulfate
concentration; 4.0, 5.5, or 7.0 pH; and primary (n) or tertiary (t)-butanol solvent isomer) were varied to determine the optimal
recovery of serologically and cellularly immunoreactive extracted proteins. A TPP extraction made with 30% ammonium sulfate and
an initial pH of 4.0 gave the best humoral and cellular immunoreactivity of caseous lymphadenitis infected goats. In particular, two immunogenic secreted (16 and 125 kDa) proteins, which had not been found by other extraction methods, were identified.