RESUMEN
A new high-performance size-exclusion chromatography method has been developed for the determination of potency of human growth hormone products. This method has been extensively validated and shown to correlate well with the hypophysectomized rat bioassay which has been used traditionally. The method is much more precise than the traditional bioassay and thus provides more reliable means of producing consistent biosynthetic human growth hormone batches.
Asunto(s)
Hormona del Crecimiento/aislamiento & purificación , Animales , Peso Corporal/efectos de los fármacos , Cartílago/efectos de los fármacos , Cartílago/crecimiento & desarrollo , Cromatografía en Gel , Cromatografía Líquida de Alta Presión , Hormona del Crecimiento/farmacología , Humanos , Indicadores y Reactivos , Ratas , Estándares de Referencia , Espectrofotometría UltravioletaRESUMEN
Two derivatives of biosynthetic human growth hormone, a sulfoxide and a mixture of two monodesamido isomers, have been isolated and characterized. The sulfoxide derivative arises from an oxidation of Met-14. The major site of deamidation is at Asn-149 with a minor site at Asn-152. In addition, a fraction has been isolated from a sample of human growth hormone that was maintained at 40 degrees C for 2 weeks. This fraction, the isolated impurities fraction, contains the sulfoxide and the desamido forms, thereby demonstrating that these derivatives are the primary degradation products of biosynthetic human growth hormone. The sulfoxide, the desamido, and the isolated impurities fraction exhibit full biological activity.