Development of fast and simple methods for porcine haptoglobin and ceruloplasmin purification

• Autores: Ana María Gutiérrez Montes, Pablo Fuentes Pardo, Laura Soler Vasco, José Joaquín Cerón Madrigal, Silvia Martínez Subiela
• Localización: Anales de veterinaria de Murcia, ISSN 0213-5434, ISSN-e 1989-1784, Nº. 26, 2010, págs. 43-54
• Idioma: español
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• 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.