Open AccessPurification of the T lymphocyte growth factor interleukin‐2 from culture media of human peripheral blood leukocytes (buffy coats)
Author(s) -
KNIEP Eva M.,
KNIEP Bernhard,
GROTE Wilhelm,
CONRADT Harald S.,
MONNER David A.,
MÜHLRADT Peter F.
Publication year - 1984
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1984.tb08359.x
Subject(s) - buffy coat , size exclusion chromatography , chromatography , chemistry , sodium dodecyl sulfate , polyacrylamide gel electrophoresis , interleukin 2 , microbiology and biotechnology , peripheral blood , lymphocyte , biochemistry , biology , in vitro , immunology , enzyme
Interleukin 2 was purified 1‐fold to apparent homogeneity from the supernatants of mitogen‐stimulated human blood leukocytes. A sequence of three purification steps was used: affinity chromatography on the bound dye cibacron blue, gel filtration on Ultrogel AcA44, and reversed‐phase high‐performance liquid chromatography on hexyl phase. The resulting interleukin 2 had a specific activity of 2 × 10 6 U/mg protein, and was free of pyrogenicity in the rabbit test. The final purification product showed two bands in sodium dodecyl sulfate/polyacrylamide gels with apparent molecular masses of 15 kDa and 17 kDa respectively. Both bands were biologically active.