Methods of selecting internalizing antibodies

Details Methods of selecting internalizing antibodies Methods of selecting internalizing antibodies

C - Chemistry, Metallurgy

40

B

C40B 40/08 (2006.01) A61K 38/00 (2006.01) A61K 47/48 (2006.01) C07K 14/01 (2006.01) C07K 16/00 (2006.01) C07K 16/28 (2006.01) C07K 16/32 (2006.01) C12N 15/00 (2006.01) C12N 15/86 (2006.01) C12Q 1/00 (2006.01) C40B 30/00 (2006.01) G01N 33/543 (2006.01) G01N 33/551 (2006.01) G01N 33/566 (2006.01)

Patent

CA 2326499

This invention provides methods of selecting antibodies that are internalized into target cells. The methods generally involve contacting target cells with one or more members of an antibody phage display library, shown in the figure. The members of the phage display library are also contacted with cells of subtractive cell line. The target cells are then washed to remove the subtractive cell line cells and members of phage display library that are non- specifically bound or weakly bound to the target cells. The target cells are cultured under conditions where members of the phage display library can be internalized if bound to an internalizing marker and internalized members of the phage display library are then identified.

L'invention concerne des procédés de sélection d'anticorps d'internalisation dans des cellules cibles. Ce procédé consiste généralement à mettre en contact des cellules cibles avec un ou plusieurs éléments d'une banque d'affichage de phages anticorps, à mettre également en contact les éléments de cette banque avec des cellules de lignée cellulaire soustractive, à laver ensuite les cellules cibles, afin d'enlever les cellules de lignée cellulaire soustractive ainsi que les éléments de la banque de phages qui ne se sont pas fixés spécifiquement -ou sont faiblement fixés- sur les cellules cibles, puis à cultiver les cellules cibles dans des conditions dans lesquelles les éléments de la banque de phages peuvent être internalisés s'ils sont fixés sur un marqueur d'internalisation, et enfin à identifier les éléments internalisés de la banque de phages.

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