Purificación y caracterización de gonadotropina coriónica humana para uso diagnóstico
Abstract
En Cuba, el UMELISA HCG, producido por el Centro de Inmunoensayo, detecta la presencia de gonadotropina coriónica humana. Con el objetivo de purificar y caracterizar gonadotropina coriónica humana (hCG) para uso diagnóstico, se desarrolló un procedimiento para su obtención a partir de orina de gestantes, con una etapa de captura y una de purificación por cromatografía de inmunoafinidad, empleando un anticuerpo monoclonal anticadena β hCG. La mayor masa de hCG en el precipitado (74,27 %) se obtuvo al 50 % de saturación del (NH4)2SO4, 22 C y 3 h de reposo. La actividad específica fue 12 417,65 UI/mg. El PAGE-SDS mostró las bandas de las cadenas α y β, con movilidad similar al control. La pureza por densitometría fue 97,3 %. Por filtración en gel se obtuvo una masa molar de 40 kDa. Con la metodología empleada se obtuvo gonadotropina coriónica humana con la calidad requerida para la obtención de los reactivos biológicos del UMELISAHCG.
Palabras clave: gonadotropina coriónica, cromatografía de afinidad, inmunoensayo, orina.
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