Membrane-translocating peptides

C - Chemistry – Metallurgy – 40 – B

Patent

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Details

C40B 20/04 (2006.01) A61K 9/127 (2006.01) A61K 47/48 (2006.01) C07K 5/00 (2006.01) C07K 7/00 (2006.01) C12N 15/00 (2006.01) C12N 15/09 (2006.01) C12Q 1/68 (2006.01) C40B 30/00 (2006.01) C40B 50/06 (2006.01) G01N 33/68 (2006.01) C07K 1/00 (2006.01)

Patent

CA 2658491

A method is provided for selecting membrane-translocating peptides (MTPs) from a peptide display library that are capable of crossing or penetrating a lipid membrane. A plurality of nucleic acid constructs that encode displayed peptides are expressed, resulting in the formation of a plurality of nucleic acid-peptide complexes, each complex comprising at least one displayed peptide associated with the corresponding nucleic acid construct encoding the displayed peptide; the complexes are exposed to a population of membrane-encapsulated compartments, allowing a translocating reaction to occur; complexes that remain unassociated with the membrane are removed; optionally complexes that are associated with the membrane are removed; and internalised nucleic acid-peptide complexes are recovered. The membrane-encapsulated compartments may be artificial vesicles such as liposomes, or populations of one or more cell types.

L'invention concerne un procédé servant à sélectionner des peptides capables d'une translocation dans une membrane (MTP) à partir d'une banque de peptides exprimés qui sont capables de traverser ou de pénétrer dans une membrane lipidique. On exprime une pluralité de constructions d'acides nucléiques qui codent pour des peptides exprimés, ce qui entraîne la formation d'une pluralité de complexes acide nucléique-peptide, chaque complexe comprenant au moins un peptide exprimé associé à la construction d'acides nucléiques correspondante codant pour le peptide exprimé ; on expose les complexes à un ensemble de compartiments encapsulés dans une membrane, permettant ainsi à une réaction de translocation d'avoir lieu ; on enlève les complexes qui restent non associés avec la membrane ; facultativement on enlève les complexes qui sont associés à la membrane ; et on récupère les complexes acide nucléique-peptide internalisés. Les compartiments encapsulés dans une membrane peuvent être des vésicules artificielles telles que des liposomes ou des ensembles d'un ou plusieurs types de cellules.

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