Purification and initial characterization of human placental trophoblast interferon induced by polyriboinosinic.polyribocytidylic acid.
Human placental trophoblast interferon (tro-IFN), induced in trophoblast cultures by a superinduction procedure, was purified to a homogeneous product with retention of biological activity. The problems associated with isolation from serum-containing medium were overcome by a combination of Blue Sepharose affinity chromatography and reversed-phase HPLC (RP-HPLC) on Separon SGX C-18. This two-step purification procedure yielded tro-IFN with a specific activity of 3.4 x 10(7) international units/mg of protein. The overall recovery of interferon activity was 66.7%. The purified tro-IFN was shown to be a glycoprotein with an Mr of 24K on native and SDS-PAGE. Its antiviral activity was stable at pH 2.0 at 37 degrees C but was sensitive to heat at 56 degrees C for 1 h and was neutralized by antibodies to human IFN-beta.
Aboagye-Mathiesen
G
G
Danish Cancer Society, Department of Virus and Cancer, Aarhus, Denmark.
Tóth
F D
FD
Juhl
C
C
Nørskov-Lauritsen
N
N
Petersen
P M
PM
Ebbesen
P
P
D002460:Cell Line; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006801:Humans; D007372:Interferons / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D008970:Molecular Weight; D010920:Placenta / Q000276:immunology*; D011070:Poly I-C / Q000494:pharmacology*; D011247:Pregnancy; D014327:Trophoblasts / Q000187:drug effects / Q000276:immunology*; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects
D007372:Interferons; D011070:Poly I-C