Methods for identifying protein-protein interactions by selective transduction
AssigneeSelective Genetics, Inc.
InventorsDavid Larocca, Andrew Baird, Paul Kassner
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Abstract
A method for identifying specific protein-protein (i.e., ligand/anti-ligand) interactions that internalize and are detected by transgene expression. A ligand displaying genetic package that carries a reporter or selectable marker and presents a ligand or putative ligand on its surface is utilized to screen cells displaying known or putative anti-ligands for the ability to successfully internalize the ligand displaying genetic package.
Claims25
A method of identifying an internalizing ligand or anti-ligand of an internalizing ligand/anti-ligand pair, comprising:
- (a) contacting one or more ligand displaying genetic packages with a cell(s), wherein the ligand displaying genetic packages comprise a candidate internalizing ligand of an internalizing ligand/anti-ligand pair, wherein the ligand displaying genetic packages carry a gene encoding a detectable product which is expressed upon internalization of the ligand displaying genetic package, and wherein the cell(s) expresses an anti-ligand-receptor fusion protein on its surface;
- (b) detecting the detectable product expressed by the cell(s);
- and
- (c) recovering a nucleic acid molecule encoding the internalizing ligand and/or a nucleic acid molecule encoding the internalizing anti-ligand from the cell(s) expressing the detectable product, and
- thereby identifying the internalizing ligand and/or anti-ligand of an internalizing ligand/anti-ligand pair.
A method of identifying an internalizing ligand and/or anti-ligand of an internalizing ligand/anti-ligand pair, comprising:
- (a) contacting one or more ligand displaying genetic packages with a cell(s), wherein the ligand displaying genetic packages comprise a candidate internalizing ligand of an internalizing ligand/anti-ligand pair, wherein the ligand displaying genetic packages carry a gene encoding a detectable product which is expressed upon internalization of the ligand displaying genetic package, and wherein the cell(s) expresses an anti-ligand-receptor fusion protein on its surface;
- (b) incubating the cell(s) under selective conditions;
- and
- (c) recovering a nucleic acid molecule encoding the internalizing ligand and/or a nucleic acid molecule encoding the internalizing anti-ligand from cell(s) which grow under the selective conditions, and
- thereby identifying the internalizing ligand and/or anti-ligand of an internalizing ligand/anti-ligand pair.
Dependencies are inferred from the claim text.