GENERAL STRATEGY FOR ANTIBODY LIBRARY SCREENING

摘要

A generally applicable method for the selective covalent attachment of a reporter molecule to a replicating entity that allows one to obtain specific binders from a single round of library screening is disclosed. For example, selective biotinylation of phage particles and yeast cells displaying a binder to any given target can be achieved via application of a coupled enzyme reaction that includes a peroxidase, an oxidase and a catalase.

主权项

1. A method for the isolation of specifically labelled replicating entities, said method comprising
a) providing in a reaction mixture a collection of replicating entities displaying variants of a receptor molecule on their surface and having either a first or a second enzyme linked to said surface, b) adding to the mixture of step a) ligand molecules that are either linked to said first enzyme if said entities are linked to said second enzyme, or said ligands are linked to said second enzyme if said entities are linked to said first enzyme, c) adding to the reaction mixture a substrate for the second enzyme, the substrate being utilized by said second enzyme in an enzymatic reaction to produce a product that is utilized by said first enzyme to convert a cosubstrate of said first enzyme into a reactive marker molecule that physically links to the respective replicating entity, thereby specifically labelling the replication entity, and d) isolating the specifically labelled replicating entities.