TY - JOUR
T1 - Isolation, purification and biochemical characterization of human placental interferons by tandem high-performance affinity chromatography
AU - Aboagye-Mathiesen, George
AU - Dalsgaard, Anne Mette
AU - Petersen, Peter M.
AU - Ebbesen, Peter
AU - Toth, Ferenc D.
AU - Zachar, Vladimir
PY - 1992/6/1
Y1 - 1992/6/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0026877211&partnerID=8YFLogxK
U2 - 10.1080/10826069208021362
DO - 10.1080/10826069208021362
M3 - Journal article
C2 - 1377824
AN - SCOPUS:0026877211
SN - 0032-7484
VL - 22
SP - 105
EP - 121
JO - Preparative Biochemistry
JF - Preparative Biochemistry
IS - 2
ER -