A method of screening for cell colonies that express a...

G - Physics – 01 – N

Patent

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Details

IPC codes

G01N 33/68 (2006.01)

Type

Patent

Patent number

CA 2531026

Description

The present invention concerns a method of identifying a cell colony which expresses a soluble variant of a target protein which method comprises (a) subjecting the cell colony to conditions which are capable of causing lysis thereof; (b) filtering the lysate through a filter having pores which allow only soluble proteins to pass through the filter; and (c) detecting target protein which has passed through the filter where the target is not detected an the basis of its own enzymatic activity. The invention further covers the identification of a cell colony expressing a soluble variant of a membrane protein using steps (a) and (b) of the method above. A kit for use in the methods is also covered by the invention.

Cette invention concerne un procédé d'identification d'une colonie cellulaire exprimant une variante soluble d'une protéine cible, lequel procédé consiste : (a) à soumettre la colonie cellulaire à des conditions pouvant provoquer la lyse de celle-ci ; (b) à filtrer le lysat à travers un filtre comprenant des pores permettant uniquement à des protéines solubles de traverser le filtre ; et (c) à détecter la protéine cible qui a traversé le filtre, la protéine cible n'étant pas détectée sur la base de sa propre activité enzymatique. L'invention concerne également l'identification d'une colonie cellulaire exprimant une variante soluble d'une protéine membranaire à l'aide des étapes (a) et (b) du procédé décrit plus haut. Cette invention concerne enfin une trousse utilisée dans les procédés de cette invention.

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