Otro
Determinação eletroanalítica de corante reativo presente como contaminante em proteínas purificadas por cromatografia de afinidade
Registro en:
Química Nova. Sociedade Brasileira de Química, v. 27, n. 3, p. 417-420, 2004.
0100-4042
10.1590/S0100-40422004000300011
S0100-40422004000300011
S0100-40422004000300011.pdf
0000-0002-2296-1393
Autor
Osugi, Marly E.
Guaratini, Claudia C. I.
Stradiotto, Nelson Ramos
Zanoni, Maria Valnice Boldrin
Resumen
Procion Green HE-4BD is a reactive dye currently used in affinity purification, and commonly present as a contaminant in the final biological preparation. An assay method is described to determine trace amounts of the dye in the presence of human serum albumin(HSA) and leakage from agarose as affinity sorbent by cathodic stripping voltammetry. The proposed method is based on the reductive peak at -0.55V in B-R buffer pH 3 (E=0V and t= 240s), obtained when samples of HSA 2% (m/v) containing dye concentrations in sodium hydroxide pH 12 are submitted to a heating time of 330 min at 80 ºC. Linear calibration curves can be obtained for RG19 dye concentrations from 5x10-9 mol L-1 to 8 x10-8 mol L-1. The detection limit (3sigma) is 1x10-9 mol L-1. Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)