Isolation, purification and biochemical characterization of human placental interferons by tandem high-performance affinity chromatography

George Aboagye-Mathiesen, Anne Mette Dalsgaard, Peter M. Petersen, Peter Ebbesen, Ferenc D. Toth, Vladimir Zachar

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10 Citations (Scopus)

Abstract

Human placental trophoblasts, fibroblasts and the trophoblast-derived malignant cell JAR are potent producers of interferons (IFNs) when stimulated with Sendai virus. The three cell lines produced different levels and compositions of IFN-α subtypes and IFN-β. Anti-IFN globulins, Cibacron Blue F3GA and Concanavalin A were covalently immobilized on pressure-stable, macroporous polymeric matrices derivatized with vinyl sulphone (HEMA-BIO 1000 VS and HEMA 1000 VS). These supports were packed in biocompatible PEEK columns and were coupled with switching valves, to develop a tandem high-performance affinity chromatographic (HPAC) method for the isoiation, purification and biochemical characterization of the IFNs produced in Sendai virus-stimulated human placental trophoblasts, fibroblasts and trophoblast-derived malignant cell, JAR, cultures. Silver-stained SDS-PAGE and gel densitometric analysis revealed the purity of the purified proteins to be between 94 and 98%. Specific activities of the purified IFNs ranged between 0.37 - 2.76 x 108 lU/mg of protein with commulative recoveries between 90 and 92.2%. The purified IFN components exhibited.

Original languageEnglish
Pages (from-to)105-121
Number of pages17
JournalPreparative Biochemistry
Volume22
Issue number2
DOIs
Publication statusPublished - Jun 1 1992

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ASJC Scopus subject areas

  • Biochemistry
  • Genetics

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