Methods and devices for identifying agents that block binding or activation of peripheral membrane proteins are disclosed.

The disclosure provides methods for carrying out Real Time Cellular Thermal Shift Assays (RT-CETSA). Also provided are molecular constructs and protein constructs for use in such assays and devices suitable for carrying out such assays.

The present invention provides a reporter for a protein fragment complementation assay characterized in that the reporter is a fused protein comprising a first fragment, a second fragment and a protein kinase sequence section, wherein the first fragment and the second fragment are derived from different sections of the same split protein, and wherein the protein kinase sequence section intervenes between the first fragment and the second fragment and wherein the kinase sequence section comprises a kinase domain sequence and one or more regulatory sequence(s). Further the invention provides polynucleotides and cells encoding for the reporter as well as methods of conducting a protein fragment complementation assay with the reporter according to the invention.

A system and method is provided for validating the manufacturing process for the production of complex biological compositions, and particularly for providing process validation information for evaluation by a federal regulatory agency. The system and method continuously and chronologically assess the concentration of proteoforms within the biological composition as it is being produced in a fermentor. Samples from the fermentor are analyzed in a pre-selected array of analysis columns, with data generated by the columns being accumulated and evaluated, and particularly compared with data from previous stages in the production process. A continuous process validation system includes top-down and bottom-up analysis sectors, each including a plurality of different analysis columns that can be selected by the controller for a particular biological composition and a particular production process.

A detection device and method for detecting a protein-based marker, and method for manufacturing a chip are provided. The detection device includes a first cover plate, in which a liquid inlet and a liquid outlet are provided; a second cover plate attached to the first cover plate to form a chamber; and a chip in the chamber. The chip includes a glass substrate and a micro-hole array layer on a side of the glass substrate. The micro-hole array layer includes a plurality of micro-holes arranged in an array, with each of the micro-holes being a nano micro-hole. The liquid inlet and the liquid outlet are configured such that a solution containing a plurality of magnetic particles enters the chamber via the liquid inlet, flows through at least a portion of the plurality of micro-holes of the micro-hole array layer, and discharges from the liquid outlet.

Characterization of the binding dynamics at the interface between any two proteins that specifically interact plays a role in myriad biomedical applications. The methods disclosed herein provide for the high-throughput characterization of the specific interaction at the interface between two protein binding partners and the identification of functionally significant mutations of one or both protein binding partners. For example, the methods disclosed herein may be useful for epitope and paratope mapping of an antibody-antigen pair, which is useful for the discovery and development of novel therapies, vaccines, diagnostics, among other biomedical applications.

A magnetic bead is respectively modified by two functional molecular layers from the inside out, termed as the polyethylene glycol (PEG) passivation layer and the photo-affinity peptide probe layer, respectively; PEG passivation layer is introduced at the surface of a magnetic bead, forming a PEG-modified magnetic bead, and the photo-affinity peptide probe layer is a molecular layer of peptide whose N-terminal end is modified with the thiol group and the diazirine group; the PEG passivation layer on the capture magnetic bead is used to reduce non-specific interaction of protein molecules, while the photo-affinity peptide probe layer can specifically recognize and capture target proteins; the weak interaction between the photo-affinity peptide probe and target proteins is converted to covalent linkage under the UV irradiation, thus achieving specific and efficient capture and magnetic separation of interacted proteins.

The present invention relates to methods and kits for detection protein-protein interactions. In particular, the present invention relates to a method for detecting the binding between a first polypeptide (A) and a second polypeptide (B) in a cell comprising i) providing a cell that expresses (a) a polypeptide (GFP1-9) comprising an amino acid sequence having at least 90% of identity with the amino acid sequence selected from the group consisting of SEQ ID NO: 1-4 (b) a first fusion protein wherein the polypeptide (A) is fused to a polypeptide (GFP10) having an amino an amino acid sequence having at least 90% of identity with the amino acid sequence selected from the group consisting of SEQ ID NO:5-7 (c) a second fusion protein wherein the polypeptide (B) is fused to a polypeptide (GFP11) having an amino an amino acid sequence having at least 90% of identity with the amino acid sequence selected from the group consisting of SEQ ID NO: 8-9 and (d) an intrabody specific for the complex formed by the self-assembly of the first, second and third polypeptides (a), (b) and (c) ii) detecting the fluorescence wherein when the fluorescence is detected it is concluded that the polypeptide (A) binds to polypeptide (B) and wherein the fluorescence is not detected it is concluded that the polypeptides (A) does not bind to polypeptide (B).

Biomolecule arrays on a substrate are described which contain a plurality of biomolecules, such as coding nucleic acids and/or isolated polypeptides, at a plurality of discrete, isolated, locations. The arrays can be used, for example, in high throughput genomics and proteomics for specific uses including, but not limited molecular diagnostics for early detection, diagnosis, treatment, prognosis, monitoring clinical response, and protein crystallography.

A method for discriminating between Parkinson's disease and multiple system atrophy, the method comprising the steps of: (1) preparing a solution containing α-synuclein monomers having a tendency to produce rod-like aggregates and/or a solution containing α-synuclein monomers having a tendency to produce twisted aggregates; (2) adding a biological sample from a subject to the solution(s) containing the α-synuclein monomers prepared in step (1); (3) allowing the α-synuclein monomers to aggregate in the solution(s) obtained in step (2); and (4) detecting α-synuclein aggregates formed in step (3).