For efficient analysis of a protein-protein interaction, the present disclosure provides a kit for analyzing a protein-protein interaction, the kit including: a 1.sup.st expression vector including a 1.sup.st polynucleotide and a multi-cloning site, wherein, the 1.sup.st polynucleotide is operably linked to a promoter and encodes a 1.sup.st fusion protein having a 1.sup.st fluorescence protein and a 1.sup.st self-assembly protein, and the multi-cloning site is a site where a polynucleotide encoding a bait protein may be operably linked to the polynucleotide encoding the 1.sup.st fusion protein; and a 2.sup.nd expression vector including a 2.sup.nd polynucleotide and a multi-cloning site, wherein, the 2.sup.nd polynucleotide is operably linked to a promoter and encodes a 2.sup.nd fusion protein having a 2.sup.nd fluorescence protein and a 2.sup.nd self-assembly protein, and the multi-cloning site is a site where a polynucleotide encoding a prey protein may be operably linked to the polynucleotide encoding the 2.sup.nd fusion protein.

The present disclosure is based on the surprising and unexpected discovery that a ligand molecule with certain characteristics is able to bind to two protein molecules simultaneously and recruit them to form a transient or stable protein-protein interaction complex. The protein-protein interaction and other cross-domain interactions gained in this process contribute additional stabilization energy to the complex beyond the combination of the binary binding energies, and therefore, largely increase the binding potency of the ligand. Accordingly, the present disclosure provides a Protein-Protein Interaction Inducing Technology (PPIIT), which includes a method to design and identify the tripartite or bifunctional compounds and use such compounds to induce protein-protein interactions in various contexts. The present disclosure also provides a composition for the purpose of inducing protein-protein interactions.

The present invention provides a regenerated cellulose membrane (RCM) that is useful in analysis of a sample for the presence of or the amount of a target molecule present in the sample. The invention also provides a method for producing and using the same. The RCM of the invention has a plurality of RCM functional groups on its surface in which a linker is covalently attached. The linker has a distal end and a proximal end, where the proximal end comprises at least one RCM linking functional group such that the RCM linking functional group is attached to the RCM functional group. The distal end of the linker comprises a receptor linking functional group that is used to covalently attach a receptor molecule. The receptor molecule is adapted for binding to a target molecule of interest when the target molecule is present in the sample. The RCM of the invention can be used for quantitative and/or qualitative analysis of a target molecule within the sample. The present invention also provides a method for producing and using the linker.

Ebolavirus is a highly lethal filovirus that causes hemorrhagic fever in humans and non-human primates. With no approved treatments or preventatives, the development of an anti-ebolavirus therapy to protect against natural infections and potential weaponization is an urgent unmet global health need. The design, biophysical characterization, and validation of peptide mimics of the ebolavirus N-trimer (“N-trimer mimics”) are described herein.

Disclosed is a library consisting essentially of a plurality of antigen-binding molecules differing in sequence from each other, wherein an antigen-binding domain in each of the antigen-binding molecules comprises at least one amino acid residue that changes the antigen-binding activity of the antigen-binding molecule depending on ion concentration conditions. Also disclosed are a composition comprising a plurality of polynucleotide molecules each encoding the antigen-binding molecules, a composition comprising a plurality of vectors each comprising the polynucleotide molecules, a method for selecting the antigen-binding molecules, a method for isolating the polynucleotide molecules, a method for producing the antigen-binding molecules, and a pharmaceutical composition comprising any of the antigen-binding molecules.

In some aspects, the invention provides methods of identifying, detecting, and/or measuring protein-protein interactions. In some aspects, the invention provides methods of identifying and/or characterizing modulators of protein-protein interactions. In some aspects, the invention provides methods of identifying and/or characterizing modulators of protein activity, wherein the methods are based at least in part on measuring interaction between a chaperone and client protein. In some aspects, the invention provides methods for identifying and/or characterizing compounds and/or for assessing compound specificity, wherein the methods are based at least in part on measuring interaction between a chaperone and client protein. In some embodiments, a client protein is a kinase. In some embodiments, a compound is a kinase inhibitor. In some aspects, the invention provides methods of profiling kinase inhibitor specificity. In some aspects, the invention provides assay systems and/or reagents useful for performing one or more of the inventive methods. In some aspects, the invention provides newly identified targets of a variety of kinase inhibitors. In some aspect, the invention provides methods of inhibiting kinases identified herein as targets of certain kinase inhibitors. In some aspects, the invention provides methods of treating a disease, e.g., cancer, by inhibiting one or more kinase(s) newly identified as targets of certain kinase inhibitors.

The present invention provides compositions and methods of use in investigations of the formation of multiprotein assemblies implicated in disease. Also provided are assays for screening candidate compounds of potential utility in preventing and/or treating such diseases by preventing the assembly of or disrupting the function of multiprotein assemblies.

A method for detecting an interaction between one or more protein bait and one or more candidate prey in a eukaryotic cell, comprising the steps of: a) providing an eukaryotic cell expressing (i) one or more protein bait, and (ii) one or more candidate prey, wherein said protein bait comprises a bait moiety and a polymerized-tubulin binding moiety. b) determining the occurence of an interaction between said one or more protein bait and said one or more candidate prey in the eukaryotic cell, wherein said protein bait is bound to polymerized tubulin in the eukaryotic cell, thereby localizing said one or more candidate prey along said polymerized tubulin, thereby detecting said interaction.

Methods for Identifying protein modulators of eukaryotic cells by expressing a combinatorial library of potential agonists inside a eukaryotic cell and then directly selecting for an agonist of a target molecule. Some methods involve co-culturing a cell expressing a combinatorial library of potential agonists and a second cell, and then selecting agents that modulate a phenotype of or a desired cellular response in the second cell. Preferably, the agonists are antibodies introduced into and expressed in the starting cells, such as agonist anti-EpoR, anti-TpoR, or G-CSFR antibodies. Also disclosed are methods for selecting from combinatorial antibody libraries bispecific antibodies that can regulate cell phenotypes.

The present disclosure relates to method and compositions for generating proteins. In particular, the present disclosure relates to electroporation mediated gene delivery in the generation of recombinant proteins (e.g., drug metabolizing enzyme and transporter vesicles) in mammalian cells.