[Article in Spanish]
Barroso P, Murcia H, Vega N, Pérez G.
Biomedica. 2005 Dec;25(4):496-510.
Laboratorio de Bioquímica, Departamento de Química, Universidad Nacional de Colombia, Bogotá, DC, Colombia.
INTRODUCTION: Egg yolk immunoglobulins (IgY) have been extensively used in immunology, biochemistry, and biotechnology in studies of human and animal health. However, their use requires two preparatory steps: first, the egg yolk lipids must be removed without impairing the immunoglobin functional properties, and second, the isolation methods must allow high recoveries for further use as detection and purification tools. OBJECTIVE: Because Tn antigen presence serves as a tumoral marker and because S. bogotensis lectin's can specifically recognize this antigen, the current study aims at making available an anti-lectin IgY. This tool will be useful in histochemical and cellular studies involving transformed cells. MATERIALS AND METHODS: Anti-lectin IgY was produced by immunization of hens with S. bogotensis lectin, and the effect of antigen dose on IgY levels was assesed. Several methods for lipid removal, IgY extraction and purification were assayed, and yields and purity of IgYs were established for each method. RESULTS: The best delipidation and extraction method included yolk dilution with water under acidic conditions and (NH4)2SO4 60% s precipitation from which 43 mg protein/yolk were recovered. Among the chromatographic methods, thiophilic chromatography permitted the recovery of a substantial quantity of pure IgY (10.4 mg IgY/yolk). With this method, the function and characteristics of IgY were preserved. CONCLUSION: The best conditions for anti-S. bogotensis functional IgY extraction and purification were established.