33 results about "Intact protein" patented technology

The invention discloses an application of knockout of PEP4 and BAR1 genes in reduction of degradation of Kluyveromyces marxianus on lactoferrin. The invention provides application of inhibiting expression of PEP4 protein and BAR1 protein in any one of the following aspects: (A1) reducing degradation of Kluyveromyces marxianus on lactoferrin; and (A2) reducing the degradation of the fermentation supernatant of the kluyveromyces marxianus on lactoferrin. Experiments prove that after the encoding gene of the PEP4 protein and the encoding gene of the BAR1 protein are knocked out in yeast cells, the lactoferrin degradation amount can be reduced, the complete protein amount is remarkably increased, and the method has important significance on increasing the secretory expression yield of the lactoferrin in kluyveromyces marxianus.

[Problems] The purpose of the present technology is to accurately detect cognitive impairment or risk thereof.[Solution] The present technology provides a cognitive impairment determination system comprising an information processing device that executes a determination step of determining presence or absence cognitive impairment or risk of cognitive impairment, based on amounts of the following biomarkers (a), (b), (c), (d), and (e) contained in a biological sample: (a) a biomarker consisting of an intact protein of Apolipoprotein A1 comprising an amino acid sequence represented by SEQ ID NO: 1, or a partial peptide thereof, (b) a biomarker consisting of an intact protein of Transthyretin comprising an amino acid sequence represented by SEQ ID NO: 2, or a partial peptide thereof, (c) a biomarker consisting of an intact protein of Complement C3 having an amino acid sequence represented by SEQ ID NO: 3, or a partial peptide thereof, (d) a biomarker Aβ1-40 consisting of a peptide having an amino acid sequence represented by SEQ ID NO: 4, and (e) a biomarker Aβ1-42 consisting of a peptide having an amino acid sequence represented by SEQ ID NO: 5.

Prokaryotic expression of a cat allergen protein chimeric peptide cFel d 1 based on a T-cell antigen epitope, and preparation of an egg yolk antibody. A chimeric peptide cFel d 1, comprising a 25-41th-position amino acid residue segment, a 46-59th-position amino acid residue segment, a 85-98th-position amino acid residue segment, a 132-140th-position amino acid residue segment and a 145-153th-position amino acid residue segment of a cat allergen Fel d 1 antigen amino acid residue. Experiments prove that an antibody obtained after immunizing animals by means of the chimeric peptide cFel d 1 has high titer, can specifically bind to a cat allergen complete Fel d 1 protein, and is superior to a polyclonal antibody prepared by means of a complete Fel d 1 protein (the titer is 8 times higher).

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 invention relates to a kit for quantitative detection of IGF-1 (Insulin-like Growth Factor-1) and a preparation and detection method thereof. The kit comprises a calibrator, a quality control product, an internal standard solution, an immune enrichment solution, a dissociation solution, a leacheate, an eluent, a mobile phase 1 and a mobile phase 2. The complete IGF-1 protein can be fully automatically detected from top to bottom, and the sample treatment efficiency is remarkably improved; wherein the matrix liquid used for preparing the kit adopts human serum without endogenous IGF-1, so that the in-vivo environment can be simulated to the greatest extent, and the detection deviation caused by different matrixes is reduced; an eluent of the kit adopts a formic acid-hydrochloric acid mixed acid-acetonitrile system, the system can provide a mild ionization environment through formic acid, and hydrochloric acid synergistically stabilizes the synergistic effect of the pH value of the solution, so that the ion inhibition effect is reduced, the response signal of IGF-1 is improved, and the signal-to-noise ratio of detection is further improved.

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.

The method comprises providing a culture of human pluripotent stem cells affixed to a first matrix comprising a first laminin; exposing the stem cells to a differential medium for a first period of time from 15 days to 50 days to obtain a first adherent cell population comprising RPE cells and / or ancestors thereof; dissociating the first population from the first matrix at the end of the first time period; re-planking the dissociated first cell population on a second substrate comprising a second laminin; and culturing the re-planked first cell population on a second substrate for a second period of time to obtain an expanded and mature second cell population comprising RPE cells. The first and second laminins may be independently selected from the group consisting of LN-521, LN-511, LN-111, and LN-121, and are intact proteins or protein fragments. In some embodiments, the cell surface marker, which can be used to generate RPE cells in vitro, is selected from the group consisting of CD140b, CD56, CD104, CD164, CD220, EGFR, GD2, CD184, CD10, CD30, CD49a, CD49b, CD50, CD171, TRA-1-60, and CD326, preferably CD140b, CD56, GD2, and / or CD184. Also included are RPE cells so produced, as well as materials and compositions that utilize such RPE cells for various treatments.

It is advantageous to employs infrared microscopy to acquire hyperspectral images of a fixed sample volume of only 2 pL of the intact protein in solution. The spectral data obtained from these images are then analyzed by correlation algorithms, specifically: two trace two-dimensions (2T2D) and two-dimensional correlation spectroscopy (2D-COS). providing both a simple output for interpretation based the 2T2D, but also the dynamic fingerprint of the desired therapeutic protein using the 2D-COS, the combination of which provide the elements for decision making.

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 provides application of SPTBN1 as a target spot in treatment of sensorineural hearing loss related to GJB2, and belongs to the technical field of sensorineural hearing. Comprising the following steps: constructing a stably transfected cell line: culturing HEK293T cells in a humidifying incubator with temperature of 37 DEG C, 95% of air and 5% of CO2 by using a DMEM (Dulbecco Modified Eagle Medium) culture medium added with 10% of fetal calf serum and 1% of double antibodies, and performing cell passage when the cells grow to a confluence degree of 80-90%; performing IP-MS analysis: purifying a protein compound by using ProteinA / G immunoprecipitation magnetic beads, taking part of extracted protein as input, then adding 2 [mu] ganti-Cx26 antibody into the residual protein extract, slightly blowing, beating and mixing, and performing turnover shaking table incubation at 4 DEG C overnight; carrying out immunofluorescence observation on co-localization of the Cx26 and the SPTBN1; constructing a stably transfected cell line for expressing the WT-Cx26 and the Mut-Cx26; and carrying out co-immunoprecipitation (Co-IP) verification: constructing a stably transfected cell line for expressing the WT-Cx26. According to the invention, limitations of treatment modes such as relatively low complete protein targeting and short treatment time window are supplemented through gene therapy, and a new insight, a new target spot and a new treatment are provided for GJB2-related hearing loss.

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.

Biomarkers for detecting cognitive dysfunction diseases and methods for detecting cognitive dysfunction diseases using the biomarkers are provided. A method for detecting cognitive dysfunction diseases comprising measuring one or more biomarkers for detecting cognitive dysfunction diseases selected from the following (a), (b), and (c) in a biological sample simultaneously or separately: (a) a biomarker for detecting cognitive dysfunction diseases consisting of an intact protein of apolipoprotein A1 comprising the amino acid sequence represented by SEQ ID NO:1 or a partial peptide thereof; (b) a biomarker for detecting cognitive dysfunction diseases consisting of an intact protein of transthyretin comprising the amino acid sequence represented by SEQ ID NO:2 or a partial peptide thereof; and (c) a biomarker for detecting cognitive dysfunction diseases consisting of an intact protein of complement C3 comprising the amino acid sequence represented by SEQ ID NO:3 or a partial peptide thereof.

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.

Provided in the present invention are a method for constructing a strain for producing a recombinant protein containing unnatural amino acids, and the strain obtained therefrom. The method comprises modifying the expression of a prfA gene contained in the strain to be controllable. The strain constructed by the method of the present invention can efficiently produce in intact protein containing unnatural amino acids, greatly reduce the production of a truncated protein and can also maintain high-speed growth.

The present invention refers to a method for determining intact glycated hemoglobin A (HbA1c) by mass spectrometry (MS), a kit and a diagnostic system adapted for performing the method.

The invention relates to a mass spectrum sampling integrated device for space-time synchronous analysis of intact protein and enzymolysis polypeptide and an analysis method, and belongs to the technical field of mass spectrum analysis. The device aims to solve the problem that synchronous analysis of complete protein and enzymolysis polypeptide is difficult to realize in the prior art. The device comprises a sample inlet, a first six-way valve, a second six-way valve, a four-way valve, a pancreatin column, a complete protein separation column C4 and a polypeptide separation column C18a. A first passage consisting of a sample inlet, a first six-way valve and a quantitative loop is formed; the second passage consists of a sample inlet, a second six-way valve, a pancreatin column, a capture column C18, a four-way valve and a polypeptide separation column C18a; and the third passage consists of a mobile phase inlet, a first six-way valve, a quantitative loop, a four-way valve and a complete protein separation column C4. The method is mainly used for realizing time-space synchronous analysis of complete protein and enzymolysis polypeptide in mass spectrometry.

It is advantageous to employs infrared microscopy to acquire hyperspectral images of a fixed sample volume of only 2 μL of the intact protein in solution. The spectral data obtained from these images are then analyzed by correlation algorithms, specifically: two trace two-dimensions (2T2D) and two-dimensional correlation spectroscopy (2D-COS). providing both a simple output for interpretation based the 2T2D, but also the dynamic fingerprint of the desired therapeutic protein using the 2D-COS, the combination of which provide the elements for decision making.

The present invention provides a system for efficiently selecting a combination of multivalent tumor neoantigens, which is intended to input multiple tumor neoantigen peptide sequences and screen out a combination with the most optimized protein sequence from a random combination peptide pool composed of these sequences. The system includes three key modules: a library construction module, an optimal path selection module, and a linker addition module. The library construction module is responsible for generating a library containing all possible combinations of input peptide sequences and calculating the potential utility of each combination. The optimal path selection module is further responsible for screening out the best protein sequence combination from these combinations to ensure that the association between peptides is both accurate and diverse. This module selects the best path for a multivalent tumor neoantigen vaccine sequence that can be generated with the least linkers, avoids the generation of unexpected neoantigens, and has a loose structure through precise screening. Finally, the linker addition module accurately adds linkers at the desired position, aiming to form a complete protein sequence that is stable and flexible in structure, while minimizing the use of linkers and optimizing the design of the overall sequence. Through this innovative method, the present invention generates a multivalent tumor neoantigen combination sequence that not only uses fewer linkers and has a simple structure, but also minimizes the generation of unexpected neoantigens, effectively addressing the challenges of multivalent antigen design.

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 disclosure relates to novel and improved methods of analyzing proteins, peptides and polypeptides by mass spectrometry using ion-ion reactions. More specifically the disclosure relates to improved methods for implementing the m / z selective arresting of ion-ion reactions within the ion-ion reaction cell of a mass spectrometer system during a period where ion-ion reactions are performed.

The invention belongs to the field of chemical analysis, and particularly relates to a method for quantitatively analyzing complete protein based on a mass spectrum tag. The specific technical scheme comprises the following steps: marking a to-be-detected protein; separating the marked protein by using a size exclusion chromatography-mass spectrometry technology; carrying out quantitative analysis on the protein to be detected by utilizing a mass spectrum mode of combining in-source cracking with multi-reaction monitoring. According to the invention, high-sensitivity absolute quantification of complete protein from top to bottom is realized on low-cost QqQ-MS through a chemical labeling-in-source cleavage-MRM combination strategy. The method for quantitatively analyzing the protein, provided by the invention, is high in universality and good in instrument compatibility.

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.

The embodiments herein provide methods to analyze the in vitro stability of recalcitrant or membrane-bound proteins in simulated gastric fluid (SGF) comprising the proteolytic enzyme, pepsin, and in combination with a novel pepsin-trypsin assay employing state-of-the-art mass spectrometric approaches, such as LC-MS / MS, to monitor the precise degradation products. The extent of protein digestion can be evaluated by the appearance of peptic products and the disappearance of tryptic peptide products (as a proxy for intact protein). The embodiments herein also provide methods for protein quantitation using high-sensitivity LC-MRM-MS quantification. The methods embodied herein are particularly useful in charactering proteins produced in transgenic plants, such as canola genetically engineered to produce long chain omega-3 polyunsaturated fatty acids.

The present invention relates to methods for detecting and quantifying intact protein-polynucleotide conjugate molecules in various sample matrices. In particular, the methods utilize triplex forming oligonucleotides in combination with protein-specific binding partners to respectively detect the polynucleotide and protein components of the conjugate molecules.

The invention discloses a protein composition used in special diet formula food for sarcopenia syndrome, a preparation method and application, and belongs to the field of food formulas for special diets. The protein composition comprises intact protein, protein hydrolysate and free amino acid, the content of leucine is 10 wt% or above, the total content of arginine and glycine is 5 wt% or above, the content of essential amino acid is 40 wt% or above, and the ratio of the intact protein to the protein hydrolysate is 80.7%: 19.3%-86%: 14%. The content of leucine, arginine and glycine in the protein composition is high, raw materials are provided for protein synthesis, the anabolic reaction of the protein is promoted, and the grip strength, the step speed and the standing-walking time of a patient suffering from the sarcopenia syndrome are improved.