12 results about "Intact protein" patented technology

The invention provides a single-chain and double-protein-expressed circular RNA construct and a preparation method and application thereof, and the single-chain and double-protein-expressed circular RNA construct comprises 5'and 3 'introns used for realizing RNA self-splicing and cyclization as a first kind of introns; the 5'and 3 'homologous arms are used as pairing sites in RNA molecules; the exon 1 and the exon 2, which are connected after being spliced, can be spliced and connected together under the action of the first type of introns to form a complete and continuous open reading frame; an interval sequence for providing spatial and adjustment sequence structures; the IRES is used for starting translation; a glycine-serine linker as a flexible linker; a green fluorescent protein for gene expression monitoring; a luciferase for producing light by a catalytic chemical reaction. All the advantages of the double-IRES system for generating natural complete protein are reserved, and the core defect of low efficiency can be thoroughly overcome.

The present invention belongs to the technical field of sensorineural hearing. Provided is the use of SPTBN1 as a target in the treatment of GJB2-related sensorineural hearing loss. The use comprises: the construction of a stably transfected cell line, wherein: HEK293T cells are cultured using a DMEM culture medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin in a humidified incubator at 37ºC with 95% air and 5% CO2, and when the cells reach 80%-90% confluence, cell passage is performed; IP-MS analysis, wherein: protein complexes are purified using Protein A / G immunoprecipitation magnetic beads, a portion of the extracted proteins is used as input, and then 2 μg of anti-Cx26 antibody is added to the remaining protein extract, same are gently pipetted and mixed, and incubated on a rotating shaker at 4℃ overnight; immunofluorescence observation of the co-localization of Cx26 and SPTBN1, wherein: a stably transfected cell line expressing WT-Cx26 and Mut-Cx26 is constructed; and co-immunoprecipitation (Co-IP) validation, wherein: a stably transfected cell line expressing WT-Cx26 is constructed. The present invention overcomes the limitations of therapeutic methods, such as the relatively low targeting specificity and short therapeutic time windows associated with full-length protein supplementation via gene therapy, thereby providing new insight into GJB2-related hearing loss, and a new target and a new treatment for same.

This invention relates to a kit for the quantitative detection of IGF-1 and its preparation and detection method. The kit includes calibrators, quality control materials, internal standard solution, immunoenrichment solution, dissociation solution, elution solution, mobile phase 1, and mobile phase 2. This invention enables fully automated top-down detection of intact IGF-1 protein, significantly improving sample processing efficiency. The matrix solution used in the kit preparation employs human serum with endogenous IGF-1 removed, maximally mimicking the in vivo environment and reducing detection bias caused by different matrix components. The elution buffer uses a formic acid-hydrochloric acid mixed acid-acetonitrile system. This system provides a mild ionization environment through formic acid, and the synergistic effect of hydrochloric acid in stabilizing the solution pH reduces ion inhibition effects, improves the IGF-1 response signal, and thus enhances the signal-to-noise ratio of the detection.

Methods comprise exposing a culture of human pluripotent stem cells adherent on a first substrate comprising a first laminin to a differentiation medium for a first time period of 15 days to 50 days to obtain a first population of adherent cells comprising RPE cells and / or progenitors thereof; at the end of the first time period, dissociating the first population from the first substrate; replating the dissociated first population of cells on a second substrate comprising a second laminin; and culturing the replated first population of cells on the second substrate for a second time period to obtain an expanded and matured second population of cells comprising the RPE cells. The first and second laminins are independently selected from LN-521, LN-511, LN-111 and LN-121, and are an intact protein or protein fragment. Also included are RPE cells and materials and compositions utilizing such RPE cells for various treatments.

The invention relates to a method for determining the ability of a molecule to modulate the activation of a G protein-coupled receptor (GPCR), said method comprising the following steps:a) introducing, in a first container:a membrane preparation bearing one or more GPCRs and one or more alpha G-proteins,a source of nonhydrolyzable or slowly hydrolyzable GTP labeled with a first member of a pair of RET partners,a ligand of the alpha subunit of a G protein (alpha G-protein) labeled with a second member of the pair of RET partners, said ligand being capable of binding to the full alpha G-protein bound to the nonhydrolyzable or slowly hydrolyzable GTP labeled with the first member of a pair of RET partners,optionally a GPCR agonist;b) measuring the RET signal emitted in the first container;c) introducing (i) in a second container, the same reagents as in step a) and the molecule to be assayed or (ii) in the first container, the molecule to be assayed;d) measuring the RET signal emitted in the second container or in the first container obtained in step c);e) comparing the signals obtained in steps b) and d), a modulation of the signal obtained in step d) relative to that obtained in step b) indicating that the molecule to be tested is capable of modulating the activation of the GPCR.

The present invention concerns a powdered nutritional composition comprising intact protein and partially hydrolysed whey protein and a dry-blending process for obtaining the powdered nutritional composition.

The invention discloses a B cell epitope peptide based on human papilloma virus E6 protein and application of the B cell epitope peptide in cancer diagnosis. According to the invention, the epitope peptide is displayed and expressed on the surface of an inert carrier bacterium S9H, so that an RI5 epitope peptide-antibody direct mediated agglutination test detection system based on thallus surface epitope display is successfully constructed. Compared with an immunoserological detection technology based on a complete protein antigen, the kit provided by the invention can be used for more specifically, sensitively, conveniently and quickly detecting the virus oncoprotein specific antibody in the body of an HPV persistent infection patient, and can be used for qualitatively and quantitatively determining the content of the antibody. The epitope peptide-antibody direct mediation agglutination test detection technology provides an efficient and reliable new method for accurate diagnosis of dynamic changes of virus oncoprotein antibodies in persistent infection and cancer stages of HPV patients, and has potential important clinical application value.

The present invention relates to a powdered nutritional composition comprising intact protein and partially hydrolyzed whey protein and a dry blending process for obtaining the powdered nutritional composition.

The present invention relates to an instant foamable beverage creamer comprising vegetable intact protein, for instance oat and / or rice protein, vegetable oil or fat, carbohydrates, and a hydrolyzed plant protein with a defined degree of hydrolysis which provides an excellent foam layer on a beverage.

The application belongs to the field of chemical analysis, and particularly relates to a method for quantitatively analyzing intact proteins based on mass spectrometry tags. The specific technical scheme comprises the following steps: labeling a protein to be measured; separating the labeled protein by using a size exclusion chromatography-mass spectrometry combined technology; and quantitatively analyzing the protein to be measured by using a mass spectrometry mode of in-source fragmentation combined with multiple reaction monitoring. The application realizes the top-down high-sensitivity absolute quantification of intact proteins on a low-cost QqQ-MS by using a chemical labeling-in-source fragmentation-MRM combined strategy. The method for quantitatively analyzing proteins provided by the application has strong universality and good instrument compatibility.

The invention relates to the technical field of gene engineering, in particular to application of a CmESI gene in regulating and controlling dwarfing of a muskmelon plant. According to the invention, methods of genetic analysis, mixed pool sequencing, Mutmap and Kasp genotype analysis and the like are combined to locate a gene causing extreme dwarfing of a plant, an SNP variation site of G / A existing in a first basic group of a seventh exon of a CmESI gene in a mutant esi is found out, the variation site causes CmESI to generate a variable shearing form with a retained sixth intron, and the mutation site is used for detecting the extreme dwarfing of the plant. A termination codon is formed in advance and cannot be translated into complete protein, so that the CmESI gene loses functions; the expression of CmESI in the 13C germplasm is inhibited by using a virus-induced gene silencing technology, and the plant height of a TRSV-CmESI plant is obviously lower than that of a control group plant; therefore, the muskmelon CmESI gene has certain theoretical significance and practical value in regulating and controlling the height of muskmelon plants.

This invention provides a mass spectrometry method for determining the 42 / 40 ratio of β-amyloid protein in human plasma, including using¹ 5 Using intact Aβ42 and Aβ40 proteins labeled with N stable isotopes as internal standards, plasma samples were extracted and purified using immunoprecipitation. Characteristic peptides were then generated through Lys-N proteolysis, and finally, targeted quantitative analysis was performed using liquid chromatography-tandem mass spectrometry. The characteristic peptide of Aβ42 is Aβ28-42 KGAIIGLMVGGVVIA; the characteristic peptide of Aβ40 is Aβ28-40 KGAIIGLMVGGVV. Compared to intact proteins, the characteristic peptides in this invention are less prone to aggregation and exhibit higher ionization efficiency in mass spectrometry, demonstrating excellent specificity, repeatability, and sensitivity, providing a reliable basis for the diagnosis of Alzheimer's disease.