The present invention provides a fibronectin type III (Fn3) molecule, wherein the Fn3 contains a stabilizing mutation. The present invention also provides Fn3 polypeptide monobodies, nucleic acid molecules encoding monobodies, and variegated nucleic acid libraries encoding such monobodies. Also provided are methods of preparing a Fn3 polypeptide monobody, and kits to perform the methods.

An apparatus for characterising a biomolecule is provided. The apparatus comprising a sample inlet channel configured to introduce a sample fluid including the biomolecule to the apparatus; an auxiliary inlet channel configured to introduce an auxiliary fluid to the apparatus; a distribution channel in fluid communication with the sample inlet channel and the auxiliary inlet channel; wherein the distribution channel is adapted to generate a distribution of biomolecules; a measurement module configured to detect a signature profile of the biomolecule to obtain a measured dataset of the detected biomolecule; a storage location configured to store and maintain a stored dataset comprising a plurality of parameters that are associated with the measured dataset obtained from the measurement module; and an analysis module configured to receive the stored dataset from the storage location and correlate the stored dataset with the measured dataset from the measurement module to provide a correlation value, wherein the analysis module is further configured to use the correlation value to determine at least two characteristics of the biomolecule simultaneously using Bayesian analysis. A method for characterising a biomolecule is also provided.

Provided herein are high-throughput, population-wide serotyping and antibody profiling assays. Disclosed variants of a Digital Serotyping assay employ next generation sequencing to measure the “serotyping profile” of barcoded subject serum antibodies tested against a range of DNA-tagged pathogen-derived antigens. The disclosed assay setup enables multiplexing in both the sample and antigen dimensions, generating a large multi-dimensional serotyping data set for more comprehensive serotyping profiling of large populations across a large number of antigens and possible pathogens. Moreover, the ability to easily scale and multiplex the number of peptide epitopes allows rapid updating of the assay content to monitor the ever-changing spectrum of pathogens. Additional applications of this technology include cancer immunology and autoimmune conditions (e.g., neoantigen or autoimmune profiling), screening for toxins, antibody therapeutics development, biosecurity, and veterinary medicine.

In certain embodiments, this disclosure relates to methods of identifying cancer agents and treating cancer with identified agents. In certain embodiments, the caner agents are capable of inducing or stabilizing SMAD4 oligomerization.

Bead arrays suitable for analysis by mass spectrometry are disclosed. In an embodiment, a bead array includes multiple reactive sites, each of the reactive sites being capable of binding multiple distinct target analytes.

A technique, called the Coupling Assay for T-cell Specificity (CATS), to identify antigen-specific cells using cell lines expressing MHC II molecules with tethered peptides. CATS successfully identified antigen-specific T cells with a low-affinity peptide, while tetramer failed to identify cells with this same peptide. Increasing avidity on artificial antigen presenting cells can overcome low affinity TCR-pMHC interactions, can identify more responding endogenous populations, and may be specific for the MHCII.

Provided herein are methods for identifying pairs of protein binding partners, mutations of which may inform the discovery of pharmaceutically useful small molecules. The methods disclosed herein may allow for the adaptation of the native protein degradation system to modulate specific disease targets at the protein level, in particular, for targets that have long been considered undruggable.

High-throughput methods for screening large populations of variant proteins are provided. The methods utilize large-scale arrays of microcapillaries, where each microcapillary comprises a solution containing a variant protein, an immobilized target molecule, and a reporter element. Immobilized target molecules may include any molecule of interest, including proteins, nucleic acids, carbohydrates, and other biomolecules. The association of a variant protein with a molecular target is assessed by measuring a signal from the reporter element. The contents of microcapillaries identified in the assays as containing variant proteins of interest can be isolated, and cells expressing the variant proteins of interest can be characterized. Also provided are systems for performing the disclosed screening methods.

The present invention provides non-natural molecules which comprise a peptide part able to stop the amyloid aggregation which is fused to a moiety which stimulates the proteasomal degradation pathway in the cell. Non-natural molecules of the invention are useful to treat human and veterinary pathological aggregation disorders.

Provided herein is synthesis of a novel acidic acid residue targeting sulfoxide-containing MS-cleavable homobifunctional cross-linker. The novel mass spectrometry-cleavable cross-linking agents can be used in mass spectrometry to facilitate structural analysis of intra-protein interactions in proteins and inter-protein interactions in protein complexes. Also disclosed herein are data based on the novel MS-cleavable homobifunctional cross-linker that are complimentary to amine-reactive sulfoxide-containing MS-cleavable reagents.