Development of fast and simple methods for porcine haptoglobin and ceruloplasmin purification
- Gutiérrez Montes, Ana María
- Fuentes Pardo, Pablo
- Soler Vasco, Laura
- Cerón Madrigal, José Joaquín
- Martínez Subiela, Silvia
ISSN: 0213-5434, 1989-1784
Año de publicación: 2010
Número: 26
Páginas: 43-54
Tipo: Artículo
Otras publicaciones en: Anales de veterinaria de Murcia
Resumen
Two fast and simple methods for porcine haptoglobin (Hp) and ceruloplasmin (Cp) purification are described in this paper. Hp was purified by ammonium sulphate fractionation and a FPLC Superdex-200 gel chromatography. The protein obtained showed two bands with a Mr (molecular mass) of about 44 kDa and 12.9 kDa corresponding to heavy (ß) and light (a) chains of Hp respectively, on sodium dodecyl sulphate-polyacrilamide gel electrophoresis, under reducing conditions. Ceruloplasmin (Cp) was isolated by one step of chromatography on amino-ethyl-derivatized Sepharose followed by gel filtration on a Superdex-200 column. The Mr of the protein, as estimated by SDS-PAGE was approximately 150 kDa. In conclusion, two new protocols have been developed for porcine Hp and Cp purification, being less time-consuming and technically demanding than those previously reported. This paper could represent an interesting guideline and be of help to obtain pure protein to use as specie-specific standard material and to produce specific antibodies.