The invention belongs to the field of chemical analysis and detection, and specifically relates to a pretreatment method for LC-MS detecting metabolomics of Aspergillus flavus. The method includes: culturing a strain of Aspergillus flavus; quenching the Aspergillus flavus; disrupting the cell membrane of Aspergillus flavus, and extracting a metabolome. The invention adopts a cold glycerol buffer solution combined with a rapid filtration method for quenching, and a MeOH/DCM/ACN/EA/HCOOH mixture is used as an metabolome extract, thereby achieving the object of efficiently extracting different polar compounds, and metabolome compound coverage is high; pretreatment of the cell metabolomics of Aspergillus flavus by the method of the invention can ensure the repeatability and stability of the metabolomics analysis method and reduce the false positive of the test results.

The present invention provides photo-cleavable anionic surfactants, particularly 4-hexylphenylazosulfonate (Azo) and sodium 4-hexylphenylazosulfonate derivatives, which can be rapidly degraded upon UV irradiation, for top-down and bottom-up proteomics. These surfactants can effectively solubilize proteins and peptide fragments with performance comparable to sodium dodecyl sulfate (SDS) and are compatible with mass spectrometry analysis of the solubilized proteins and peptide fragments. Top-down proteomic studies using the present photo-cleavable anionic surfactants has allowed the detection of 100-fold more unique proteoforms as compared to controls and has enabled the solubilization of membrane proteins for comprehensive characterization of protein post-trans-modifications. In addition, the present photo-cleavable anionic surfactants are also suitable for dissolving polypeptides in bottom-up proteomic experiments including extracellular matrix proteomics, and are suitable as a substitute for SDS in gel electrophoresis.

The disclosure provides a lipid nanodisc including a lipid bilayer having two opposing hydrophilic faces and a hydrophobic edge between the hydrophilic faces, and a copolymer encircling the hydrophobic edge of the lipid bilayer, the copolymer including a first monomeric unit including a pendant aromatic group, and a second monomeric unit including a pendant hydrophilic group, wherein the first monomeric unit and the second monomeric unit are present in the copolymer is a molar ratio ranging from 1:1 to 3:1 for the first monomeric unit:the second monomeric unit. The disclosure further provides a method of making the polymer-based lipid nanodiscs of the disclosure and methods of characterizing membrane proteins using the polymer-based lipid nanodiscs of the disclosure.

Methods and systems for identifying a protein within a sample are provided herein. A panel of antibodies are acquired, none of which are specific for a single protein or family of proteins. Additionally, the binding properties of the antibodies in the panel are determined. Further, the protein is iteratively exposed to a panel of antibodies. Additionally, a set of antibodies which bind the protein are determined. The identity of the protein is determined using one or more deconvolution methods based on the known binding properties of the antibodies to match the set of antibodies to a sequence of a protein.

Disclosed are methods of quantifying multiple quality attributes, such as post translational modifications, of multiple samples in a single mass spectrometry (MS) run, including contacting two or more samples with a digesting solution under conditions sufficient to digest samples, wherein each sample is digested separately and the digesting solution is a Tris-free buffer solution; contacting each of the two or more digested samples with a specific Tandem Mass Tag (TMT) labeling reagent under conditions sufficient to label peptides within each of the digested samples with the specific TMT labeling reagent; quenching labeling of peptides within each of the two or more digested samples; combining equal volumes of the two or more labeled, digested samples into a single combined sample solution; and analyzing the single combined sample solution by targeted mass spectral analysis, thereby allowing multiple quality attributes of the two or more samples to be quantified in a single mass spectrometry (MS) run.

The present disclosure relates to glycoproteomics. More specifically, the current disclosure provides methods for determining one or more of the glycoproteins, glycosylation sites, glycopeptide fragments, and glycan compositions of both membrane and cytosolic proteins. The methods herein employ a single processing method that enables extraction of membrane and cytosolic proteins for the identification and analysis of whole-cell glycosylation, independent of species or sample type.

The disclosure relates to methods of analyzing a post-translationally modified protein of interest using electrophoresis, the methods comprising deglycosylating the protein of interest after labeling.

The application relates to methods and systems for proteomics and spatial mapping of biomolecules using a next generation sequencing readout to decipher biomolecular and cellular interaction networks. Specifically, disclosed are antenna networks generated by conjugating DNA antennas to proteins. The antennas carry a unique antenna identifier (UAI) sequence that can provide spatial location of the network, as well as biomolecules by transfer of the UAI to reporter oligonucleotides associated with other antennas and biomolecules. The methods and systems are also applicable to single cells.

Method of activating dysfunctional T cells, determining responsiveness of a subject having a tumor to an immune checkpoint inhibition are provided. Also provided is an agent capable of inhibiting a target gene or expression product thereof for use in treating a subject having a tumor. Additionally or alternatively provided is an immune checkpoint inhibitor and an agent capable of inhibiting a target gene or expression product thereof for use in treating a subject having a tumor.

Multiplexed test measurements are conducted using an assay module having a plurality of assay domains. In preferred embodiments, these measurements are conducted in assay modules having integrated electrodes with a reader apparatus adapted to receive assay modules, induce luminescence, preferably electrode induced luminescence, in the wells or assay regions of the assay modules and measure the induced luminescence.