Selective posttranslational modification of phage-displayed...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/00 (2006.01) C12P 21/04 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2624290

The invention relates to posttranslational modification of phage-displayed polypeptides. These displayed polypeptides comprise at least one unnatural amino acid, e.g., an aryl-azide amino acid such as p-azido-L-phenylalanine, or an alkynyl-amino acid such as para- propargyloxyphenylalanine, which are incorporated into the phage-displayed fusion polypeptide at a selected position by using an in vivo orthogonal translation system comprising a suitable orthogonal aminoacyl-tRNA synthetase and a suitable orthogonal tRNA species. These unnatural amino acids advantageously provide targets for posttranslational modifications such as azide-alkyne [3+2] cycloaddition reactions and Staudinger modifications.

L'invention concerne la modification post-traduction de polypeptides exprimés à la surface des phages. Ces polypeptides comportent au moins un aminoacide artificiel, par exemple, un aminoacide arylazide tel que p-azido-L-phénylalanine, ou un aminoacide alkynyle tel que para-propargyloxyphénylalanine, lesquels sont introduits dans le polypeptide de fusion exprimé à la surface des phages dans une position sélectionnée à l'aide d'un système de traduction orthogonal in vivo qui comprend une aminoacyl-ARNt synthétase ainsi qu'une variété d'ARNt orthogonale appropriée. Ces aminoacides artificiels constituent avantageusement des cibles pour les modifications post-traduction telles que les réactions de cycloaddition azide-alkyne [3+2] et les modifications de Staudinger.

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