Preparative in vitro synthesis of bioactive human interleukin-2 in a continuous flow translation system.
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.
Kolosov
M I
MI
Research Center of Molecular Diagnostics and Therapy, Moscow, Russia.
Kolosova
I M
IM
Alakhov VYu
Ovodov SYu
Alakhov
Y B
YB
D000327:Adsorption; D000595:Amino Acid Sequence; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002474:Cell-Free System; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010084:Oxidation-Reduction; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000494:pharmacology; D014158:Transcription, Genetic
D007376:Interleukin-2; D012333:RNA, Messenger; D011994:Recombinant Proteins