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AssigneeThe Scripps Research Institute
InventorsHongkai Zhang, Ian A. Wilson, Richard A. Lerner
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The invention provides methods for identifying protein modulators (e.g., antibody agonists) of eukaryotic cells. The methods typically involve expressing a combinatorial agent library (e.g., via lentiviral vectors) inside a eukaryotic cell type (e.g., a mammalian cell) and then directly selecting for agents (e.g., antibodies) that are agonist of a target molecule (e.g., a signaling receptor) that modulates a phenotype of or elicits a cellular response in the cell. Some related methods involve co-culturing a cell expressing a combinatorial agent library and a second cell, and then selecting agents that modulate a phenotype of or elicit a cellular response in the second cell. Preferably, the agents are antibodies and are introduced into and expressed in the starting cells under conditions each individual cell expresses no more than 3 different members of the antibody library. In addition, the invention provides methods for identifying protein agonists that capable of reprograming or trans-differentiating a target cell. Also provided in the invention are specific agonist antibodies of signaling receptors or biomolecules that modulate a phenotype or effectuate a cellular response in a eukaryotic cell (e.g., agonist antibodies of EpoR, TpoR or G-CSFR). Further provided in the invention are methods for selecting from combinatorial antibody libraries bispecific antibodies that can regulate cell phenotypes.
A method for identifying a polypeptide agent capable of reprograming or trans-differentiating a target cell, comprising (1) expressing a library of secreted or membrane-integrated candidate polypeptide agents in a reporter cell bearing a signaling receptor expressed by the target cell to produce a heterogeneous population of modified, agent-expressing reporter cells, (2) identifying, in an autocrine manner, a subpopulation of agent-expressing reporter cells with activated signaling of the receptor, (3) isolating a group of agonist agents from the identified subpopulation of cells, and (4) contacting the group of agonist agents with a population of the target cell and selecting a cell with a specific phenotype indicative of trans-differentiation;
A method for identifying an antibody modulator of a eukaryotic cell type, comprising (a) expressing in a homogeneous population of cells of a second cell type a library of candidate antibodies or antigen-binding fragments thereof under conditions each cell expresses no more than 3 different antibody species to produce a heterogeneous population of antibody-expressing cells, (b) co-culturing a population of cells of said eukaryotic cell type and the population of antibody-expressing cells, and (c) selecting a specific antibody-expressing cell that alters a phenotype of said eukaryotic cell type;
An antibody or antigen-binding fragment thereof which has the same binding specificity as that of an antibody comprising (1) heavy chain CDR1, CDR2 and CDR3 sequences respectively shown in SEQ ID NO:35, SEQ ID NO:36 and SEQ ID NO:37;
Dependencies are inferred from the claim text.