CELL-FREE TRANSLATION SYSTEM FOR THE PRODUCTION OF (CHEMICALLY) MODIFIED PROTEINS

摘要

The present invention relates to a cell-free translation system comprising a nonsense-codon suppressing agent and an anti-release factor antibody which precipitates and/or crosslinks a release factor in said cell-free translation system. The present invention further relates to a kit comprising said cell free translation system. Moreover, the present invention relates to an anti-release factor antibody directed against release factor of Thermus thermophiles and, inter alia, obtainable by eliciting an in vivo humoral response against release factor 1 from Thermus thermophilus or a fragment thereof in a non-human vertebrates. Furthermore, the present invention relates to the use of an anti-release factor antibody for the preparation of a cell-free translation system. Moreover, the present invention relates to a method for the production of an anti-release factor antibody comprising the steps of eliciting an in vivo humoral response against release factor 1 from Thermus thermophilus or a fragment thereof in a non-human vertebrate. Furthermore, the present invention provides for a method for the production of alloproteins, comprising the step of translating RNA into a translation product in the cell-free translation system of the present invention. Further, the present invention relates to a puromycin derivative having a residue covalently attached directly or via a linker to one of its elements selected from the group consisting of the element N⁴ of the cytosine-residue of an 5' attached cytidine-residue and the element C⁵ of the cytosine-residue of an 5' attached cytidine-residue. Moreover, the present invention relates to the use of a puromycin derivative having a residue covalently attached directly or via a linker to one of its elements selected from the group consisting of the element N⁴ of the cytosine-residue of an 5' attached cytidine-residue and the element C⁵ of the cytosine-residue of an 5' attached cytidine-residue, in the cell-free translation system of the present invention, in the method for the production of alloproteins of the present invention. The invention also provides for specific kits described herein.