Methods to select for agents that stabilize protein complexes
First Claim
1. A method for selecting a conformation-selective binding agent of a transient protein complex comprising a conformational epitope, the method comprising the steps of:
- a) displaying a collection of binding agents at the extracellular surfaces of a population of cells; and
b) utilizing cell sorting to separate and select, from the population of cells, cells displaying binding agents thati. specifically bind to a conformational epitope of the transient protein complex and not to the individual members of the transient protein complex, and/orii. specifically bind to a conformational epitope of one of the individual members of the transient protein complex and not to a conformational epitope of the transient protein complex nor to the other individual member(s) of the transient protein complex, and/oriii. specifically bind to a conformational epitope of one of the individual members of the transient protein complex and to a conformational epitope of the transient protein complex and not to the other individual member(s) of the transient protein complex;
wherein the transient protein complex comprises at least two interacting individual members; and
wherein the individual members of the transient protein complex are distinguishably tagged.
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Abstract
The application relates to the field of structural biology. More specifically, the disclosure relates to methods for the identification and characterization of biomolecular tools allowing the selective recognition and/or stabilization of distinct conformational states of protein complexes, including transient protein-protein interactions and protein-nucleic acid complexes. Such tools can then be used for purification purposes, crystallization and structure determination of these stabilized protein complexes, for drug discovery, as research tools, as well as for diagnosis and treatment of diseases.
1 Citation
18 Claims
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1. A method for selecting a conformation-selective binding agent of a transient protein complex comprising a conformational epitope, the method comprising the steps of:
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a) displaying a collection of binding agents at the extracellular surfaces of a population of cells; and b) utilizing cell sorting to separate and select, from the population of cells, cells displaying binding agents that i. specifically bind to a conformational epitope of the transient protein complex and not to the individual members of the transient protein complex, and/or ii. specifically bind to a conformational epitope of one of the individual members of the transient protein complex and not to a conformational epitope of the transient protein complex nor to the other individual member(s) of the transient protein complex, and/or iii. specifically bind to a conformational epitope of one of the individual members of the transient protein complex and to a conformational epitope of the transient protein complex and not to the other individual member(s) of the transient protein complex; wherein the transient protein complex comprises at least two interacting individual members; and wherein the individual members of the transient protein complex are distinguishably tagged. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method for selecting a conformation-selective binding agent of a conformational epitope of a transient protein complex, the method comprising:
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a) displaying a collection of binding agents at the extracellular surfaces of a population of cells; and b) utilizing cell sorting to separate and select, from the population of cells, cells displaying binding agents that i. specifically bind to the conformational epitope of the transient protein complex and not to the individual members of the transient protein complex, and/or ii. specifically bind to one of the individual members of the transient protein complex and to the conformational epitope of the transient protein complex and not to the other individual member(s) of the transient protein complex; wherein the transient protein complex comprises at least two interacting individual members; and wherein the individual members of the transient protein complex are distinguishably tagged.
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Specification