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Preparative in vitro synthesis of bioactive human interleukin‐2 in a continuous flow translation system
Author(s) -
Kolosov MI,
Kolosova IM,
Alakhov VYu,
Ovodov SYu,
Alakhov YB
Publication year - 1992
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1992.tb00217.x
Subject(s) - translation (biology) , recombinant dna , in vitro , continuous flow , messenger rna , protein biosynthesis , chemistry , interleukin , interleukin 2 , ribonucleoprotein , biochemistry , cytokine , biology , immunology , rna , gene , physics , mechanics
Recombinant human interleukin‐2 has been synthesized in vitro in a continuous flow translation system based on the wheat germ extract. In the course of translation of mRNA the interleukin‐2 becomes aggregated due to the adsorption of this protein onto the ribonucleoprotein complex. This process correlates with the cessation of translation that is usually observed in 25–30 min. This can be prevented by the use of a flow system that allows continuous removal of the synthesized protein and maintains a steady concentration of all the necessary components. This approach permitted a yield of 1,500 protein molecules per mRNA molecule. The interleukin obtained promotes the proliferation of the interleukin‐2‐dependent CTLL‐2 cell line. The biological activity of interleukin‐2 not subjected to oxidative refolding was 10(5) units per milligram of protein.