Resumo

GUISADO-BOURZAC, Frenkel et al. Obtaining an affinity support for phospholipase A₂ purification. Rev Cub Quim [online]. 2016, vol.28, n.2, pp.595-609. ISSN 2224-5421.

The synthesis of a glyoxyl-Sepharose support was achieved departing from Sepharose CL-4B firstly activated, oxidized and aminated. Amination process is done in order to obtain an amino gel that could join specific ligands for phospholipases A₂ purification. The egg yolk phosphatidylcholine (ePC) was immobilized by covalent method and was controlled by phosphate determination in the departure material and in laundries. This support has the advantage of being able to be chemically modified achieving different affinity supports. The validity of the supports obtained was checked by the addition of chromatography fractions from the sea anemones Condylactis gigantea and Stichodactyla helianthus, and the known snake venom from Crotalus durisus terrificus with phospholipase A₂ activity. The typical elution maximum corresponding was obtained. Phospholipase A₂ activity was corroborated qualitatively by a TLC-based method after exposure to purified ePC and fluorogenic substrate 1-palmitoil-2-NBD-C12-PC

Palavras-chave : affinity chromatography; phospholipase A₂; egg yolk phosphatidylcholine immobilization; sea anemone; amine gel.

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