Purification and in vitro labeling of interferon from a human fibroblastoid cell line

Journal Article · Tue Jul 25 04:00:00 EDT 1978 · J. Biol. Chem.; (United States)

OSTI ID:7031965

Berthold, W; Tan, C; Tan, Y H

Interferon was produced from a clonal human fibroblastoid line. This cell line was derived from an established fibroblastoid culture treated with ethylmethane sulfonate and is capable of producing higher amounts human interferon than primary human fibroblast cultures. The interferon produced from this cell line was purified by concanavalin A and phenyl-Sepharose column chromatography followed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified interferon was then labeled with (/sup 125/I)-iodine, with (3H)5-dimethylaminonaphthalene-1-sulfonyl-chloride and with sodium (3H)borohydride after periodate oxidation. An appreciable amount of biological activity was retained after both tritium labelings but not after iodination. When subjected to electrophoresis in the presence of sodium dodecyl sulfate all three radioactive interferon preparations comigrated with the purified interferon preparation as a single protein component with a molecular weight of 19,000. Both purified and radioactive preparations were also shown to migrate with the antiviral activity in polyacrylamide gels in the absence of sodium dodecyl sulfate. The specific activity of purified interferon preparations ranged from 2 X 10/sup 8/ to 10 X 10/sup 8/ units/mg of protein.

• Research Organization: Univ. of Calgary, Alberta
• OSTI ID: 7031965
• Journal Information: J. Biol. Chem.; (United States), Journal Name: J. Biol. Chem.; (United States) Vol. 253:14; ISSN JBCHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DIAGNOSTIC TECHNIQUES
ELECTRON CAPTURE RADIOISOTOPES
FIBROBLASTS
HYDROGEN ISOTOPES
INTERFERON
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
LABELLED COMPOUNDS
LABELLING
LIGHT NUCLEI
MAMMALS
MAN
NUCLEI
ODD-EVEN NUCLEI
PRIMATES
PURIFICATION
RADIOISOTOPES
SOMATIC CELLS
TRITIUM
VERTEBRATES
YEARS LIVING RADIOISOTOPES