51 results about "Folding protein" patented technology

The invention relates to recombinant pichia pastoris for high expression of porcine pancreatic phospholipase A2 and application of the recombinant pichia pastoris, and belongs to the technical field of genetic engineering. According to the method disclosed by the invention, a vacuole sorting receptor Vps10 is knocked out by utilizing a CRISPR-Cas9 system, a CPY sorting way between a trans-golgi apparatus and a vacuole is interrupted, and ppPLA2 degradation is reduced; then, in order to relieve unfolded protein response in the endoplasmic reticulum, molecular chaperones are introduced into the genome, and synthesis of ppPLA2 is enhanced; the expression intensity of the ppPLA2 is improved by increasing the gene copy number of the ppPLA2; and finally, high-density fermentation is performed in a 3-L bioreactor, so that high-efficiency expression of the ppPLA2 in the pichia pastoris is realized.

The invention provides application of a biomarker combination for early screening and diagnosis of transthyretin cardiac amyloidosis (ATTR-CA) high-risk groups in preparation of a product for early screening and diagnosis of the ATTR-CA high-risk groups. In general, one or more biomarkers, such as total prealbumin (total PA), transthyretin (TTR) tetramer, misfolded protein, protein misfolding rate, and soluble growth stimulus expression gene 2 protein (sST2). In ATTR-CA high-risk groups and non-high-risk groups, the biomarkers show significant difference, which prompts that the biomarkers are related to ATTR-CA. When a plurality of biomarkers are combined for use, the biomarkers have relatively high sensitivity and specificity in early screening and diagnosis of ATTR-CA high-risk groups, have relatively high diagnosis efficiency, and provide a new thought and strategy for early diagnosis and treatment of ATTR-CA.

The present invention provides a multiomics approach, which integrate single-cell RNA-sequencing (scRNA-seq) and spatiotemporal transcriptomics (ST) offering potential for dissecting transcriptional networks and revealing cell-cell interactions involved in biomolecular pathomechanisms. The present invention also provides a multimodal approach combining high-throughput scRNA-seq and ST to elucidate XLRS-specific transcriptomic signatures in two XLRS-like models with retinal splitting phenotypes, including genetically engineered (Rs1emR209C) mice and patient-derived retinal organoids harboring the same patient-specific p.R209C mutation. Through multiomics transcriptomic analysis, the endoplasmic reticulum (ER) stress / eIF2 signaling, mTOR pathway, and the regulation of eIF4 and p70S6K pathways as chronically enriched and highly conserved disease pathways between two XLRS-like models are identified. Western blots and proteomics analysis validated the occurrence of unfolded protein responses, chronic eIF2α signaling activation, and chronic ER stress-induced apoptosis. Furthermore, therapeutic targeting of the chronic ER stress / eIF2α pathway activation synergistically enhanced the efficacy of AAV mediated RS1 gene delivery, ultimately improving bipolar cell integrity, postsynaptic transmission, disorganized retinal architecture and electrophysiological responses. Collectively, the complex transcriptomic signatures obtained from Rs1emR209C mice and patient-derived retinal organoids using the multiomics approach provide opportunities to unravel potential therapeutic targets for incurable retinal diseases, such as XLRS.

Provided herein are methods of protein synthesis, protein synthesis reagents for the expression of eukaryotic and mammalian proteins and methods for optimizing protein expression yields for mammalian proteins. The methods are applicable to protein expression on a microfluidic device having hydrophobic surfaces by merging droplets on the device in order to screen a selection of expression compositions in parallel and then identify the optimum conditions for producing the stable folded protein in a cell-based system.

The present invention provides a method for producing a refolded protein, the method comprising a step for mixing, inside a micro mixer, a buffer and a solubilization solution for a protein that has lost higher order structure or that has become insoluble.

The present invention provides compounds, prodrugs, and pharmaceutically acceptable salts thereof that directly inhibit IRE-1α activity in vitro. Such compounds and prodrugs are useful for treating diseases associated with the unfolded protein response or regulated IRE1-dependent decay (RIDD) and can be used as single agents or in combination therapies.

Antibodies can treat neurodegenerative disorders (NDs) caused by misfolded proteins. Still, the blood-brain-barrier (BBB) resists their entry, a barrier that can be reduced by scFv instead of whole antibody and conjugating it with transferrin protein to induce transcytosis. While the scFv binds variable heavy and light chains with a non-cleavable linker, a cleavable linker between the scFv and transferrin protein can reduce exocytosis, enhancing the activity of the scFv.

The application discloses a kind of short peptides simulating RhoE C end, derivative and its pharmaceutical use in treating ventricular remodeling, belong to biological medicine technical field.The short peptide and derivative contain RhoE C end 200-240 linear function domain, can be combined and activated WWP2, remove HGS self-inhibition, start endosome-lysosome degradation pathway.Short peptide sequence is as SEQ ID NO.1, derivative can be coupled with TAT, T7 or RVG membrane penetrating peptide (SEQ ID NO.2-4), and endocytosed to myocardial cell by caveolin high efficiency delivery.The application solves the defects that existing anti-ventricular remodeling drug inhibits protective autophagy flow, and the ability of eliminating toxic protein is limited, can efficiently eliminate damaged mitochondria and misfolded protein, inhibits myocardial hypertrophy, fibrosis, improves cardiac remodeling, provides safe and effective treatment strategy for related diseases.

A method for determining the presence of soluble, misfolded protein in or on the eye of a subject is described. The method includes contacting the eye of the subject with a monomeric protein to form an incubation mixture; incubating the incubation mixture to form a misfolded protein aggregate from the monomeric protein; and determining if a detectable amount of misfolded and / or aggregated protein is present in or on the eye; wherein detection of misfolded and / or aggregated protein indicates the presence of soluble, misfolded protein in the eye. Use of the method for diagnosing disease associated with protein misfolding in a subject is also described.

The application discloses a liposome vaccine of Acinetobacter baumannii outer membrane protein and a preparation method and application thereof, and relates to the field of biological medicines. The application efficiently expresses the Acinetobacter baumannii outer membrane protein through a genetic engineering method, and obtains a liposome vaccine of Acinetobacter baumannii outer membrane protein by combining the obtained various outer membrane proteins as antigens with a lipid mixture and an adjuvant. It is verified that the vaccine can provide complete protection for mice attacked by a lethal dose of Acinetobacter baumannii. In the application, the outer membrane protein loaded in the liposome is re-folded in structure, the immunogenicity of the protein is enhanced, and the liposome can reduce the toxicity of the outer membrane protein. Omp38+BauA+FimA can be an effective antigen combination for preventing Acinetobacter baumannii infection. Pam2CSK4 as a TLR2 agonist has a strong auxiliary effect on the liposome vaccine of Acinetobacter baumannii, and can induce the activation of an innate immune response, thereby providing more comprehensive protection for a host.

Antibodies can treat neurodegenerative disorders (NDs) caused by misfolded proteins. Still, the blood-brain-barrier (BBB) resists their entry, a barrier that can be reduced by scFv instead of whole antibody and conjugating it with transferrin protein to induce transcytosis. While the scFv binds variable heavy and light chains with a non-cleavable linker, a cleavable linker between the scFv and transferrin protein can reduce exocytosis, enhancing the activity of the scFv.

A number of protein aggregation diseases are associated with accumulation of misfolded proteins, which are known as protein aggregates, including, but not limited to, neurodegenerative and non-degenerative diseases and disorders. The present disclosure provides an assay, compositions and kits for the qualitative and quantitative assessment of aggregated proteins in solution using a microparticle immunocapture assay that combines the advantages inherent to a specific first and second capture moiety that binds specifically to an aggregated protein which can reveal at the same time the amount and the size of aggregates measured in a sample, fluid, tissue, cavity, or pharmacological product.

A compound specifically binding to an α-synuclein aggregate, and a preparation method therefor and the use thereof. Specifically, the compound binding to the α-synuclein aggregate comprises compounds as shown in formula A or sub-general formulas thereof, or stereoisomers, pharmaceutically acceptable salts, solvates or stable isotope variants thereof. The compound is a small molecule tracer, which can specifically recognize the α-synuclein aggregate, and can be used for the preparation of a drug for the treatment or diagnosis of neurodegenerative diseases (such as Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Alzheimer's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy and progressive muscular atrophy) related to the α-synuclein aggregate and other misfolded proteins.

Provided herein are methods of protein synthesis, protein synthesis reagents for the expression of eukaryotic and mammalian proteins and methods for optimizing protein expression yields for mammalian proteins. The methods are applicable to protein expression on a microfluidic device having hydrophobic surfaces by merging droplets on the device in order to screen a selection of expression compositions in parallel and then identify the optimum conditions for producing the stable folded protein in a cell-based system.

To provide an electrode for electrochemical measurement capable of detecting a target substance without bonding an index substance, and a measuring method.SOLUTION: A measurement method includes a binding step of bringing a specimen solution 8 containing a target substance 7 into contact with an electrode 2 in which a naturally denatured protein 22 having a naturally denatured region that is folded by specifically binding to the target substance 7 is immobilized as an identifier on a surface of a conductive layer 21 to specifically bind the target substance 7 to the naturally denatured protein 22, and a measurement step of bringing the electrode 2 brought into contact with the specimen solution 8 in the binding step into contact with an electrolyte solution 6 containing redox substances Red and Ox to perform electrochemical measurement. The presence or concentration of the target substance 7 in the specimen solution 8 is measured by detecting or quantifying an increase in a surface exposure area of the conductive layer 21 caused by folding of the naturally denatured protein 22 by bonding with the target substance 7.SELECTED DRAWING: Figure 3

Antibodies can treat neurodegenerative disorders (NDs) caused by misfolded proteins. Still, the blood-brain-barrier (BBB) resists their entry, a barrier that can be reduced by scFv instead of whole antibody and conjugating it with transferrin protein to induce transcytosis. While the scFv binds variable heavy and light chains with a non-cleavable linker, a cleavable linker between the scFv and transferrin protein can reduce exocytosis, enhancing the activity of the scFv.

The invention relates to in situ methods for detecting the presence of misfolded proteins in samples (e.g. tissue samples). The invention also relates to the use of said methods in the detection of a proteinopathy or an increased risk thereof in a subject and to methods of determining the efficacy of therapeutic interventions.

Antibodies can treat neurodegenerative disorders (NDs) caused by misfolded proteins. Still, the blood-brain-barrier (BBB) resists their entry, a barrier that can be reduced by scFv instead of whole antibody and conjugating it with transferrin protein to induce transcytosis. While the scFv binds variable heavy and light chains with a non-cleavable linker, a cleavable linker between the scFv and transferrin protein can reduce exocytosis, enhancing the activity of the scFv.

Antibodies can treat neurodegenerative disorders (NDs) caused by misfolded proteins. Still, the blood-brain-barrier (BBB) resists their entry, a barrier that can be reduced by scFv instead of whole antibody and conjugating it with transferrin protein to induce transcytosis. While the scFv binds variable heavy and light chains with a non-cleavable linker, a cleavable linker between the scFv and transferrin protein can reduce exocytosis, enhancing the activity of the scFv.

The present invention discloses a highly active unfolded protein response element and its screening method and application. The screening method can efficiently and quickly screen out UPRE elements with high response activity. The obtained highly active UPRE2 element has been improved in both response intensity and response amplitude. Tandem connection of these highly active UPRE elements can further improve their performance. Inserting the obtained highly active UPRE2 element upstream of the core region of the promoter can achieve dynamic regulation of synthetic pathway genes, improve the yield of target products such as recombinant proteins or natural products, and has broad application prospects and important practical significance.

A conjugate comprising (i) a component of PROTAC or (ii) PROTAC has been developed that enhances the efficacy of PROTAC technology by increasing selective delivery and absorption to inflammatory sites such as the brain, and a dendrimer formed from a hydroxylated polyamidoamine (hydroxyl PAMAM) or sugar, e.g., a glucose dendrimer. Since the conjugate can be used for targeted intracellular delivery to degrade misfolded proteins and / or disease-related proteins located in the cytoplasm, the conjugate can utilize the cell's intracellular protein degradation mechanism.

The present invention discloses a design method of a novel probe CazFPC-SNAP and its visualization detection of protein aggregates. The CazFPC-SNAP is synthesized by a three-step method of conjugating phenylboronic acid pinacol ester with a 4-hydroxybenzylidene imidazolinone core and an anchoring biotag SNAP. Phenylboronic acid pinacol ester serves as a hydrogen peroxide recognition site, and the introduced biotag SNAP is used to identify and label the target protein. The probe CazFPC-SNAP can not only be used to quickly detect the concentration level of hydrogen peroxide, but also can realize the visualization imaging of protein misfolding and aggregation in living cells. It has an obvious fluorescent response to the viscosity change in the protein folding system and can identify protein aggregates with the naked eye. It has low cytotoxicity, good biocompatibility, and stable fluorescent properties. It has extremely important applications in the fields of bioimaging, fluorescent labeling, biosensors, and drug development. The present invention solves the problem that insoluble misfolded proteins in cells are difficult to detect.

Antibodies can treat neurodegenerative disorders (NDs) caused by misfolded proteins. Still, the blood-brain-barrier (BBB) resists their entry, a barrier that can be reduced by scFv instead of whole antibody and conjugating it with transferrin protein to induce transcytosis. While the scFv binds variable heavy and light chains with a non-cleavable linker, a cleavable linker between the scFv and transferrin protein can reduce exocytosis, enhancing the activity of the scFv.

The invention relates to the technical field of biological medicines, in particular to application of beta-aminoisobutyric acid in preparation of a preparation for treating animal mastitis. The invention discloses application of beta-aminoisobutyric acid in preparation of a preparation for treating animal mastitis. The beta-aminoisobutyric acid provided by the invention is derived from the posterior intestine content of the healthy saanen milk goat with the lowest somatic cell count in the milk obtained by large group screening. According to the present invention, the proliferation efficiency of dairy cow mammary epithelial cells can be significantly promoted, and the expression of the related proteins involved in the endoplasmic reticulum protein quality control pathway can be promoted in the LPS-induced dairy cow mammary epithelial cell inflammation model so as to enhance the folding efficiency of the proteins during the inflammation process; the removal of mistakenly folded and unfolded proteins is accelerated, and finally, the concentration of proinflammatory cytokines is reduced, and the concentration of anti-inflammatory cytokines is increased, so that the inflammatory phenotype is relieved.

The invention provides a mu-conopeptide fusion protein and a preparation method of mu-conopeptide, and belongs to the technical field of biology. The invention provides a [mu]-conopeptide fusion protein. The [mu]-conopeptide fusion protein comprises a folded protein in a connected state and [mu]-conopeptide which is repeatedly connected in series, the folded protein comprises thioredoxin and disulfide bond isomerase; the amino acid sequence of the mu-conopeptide is as shown in SEQ ID NO: 1. According to the method disclosed by the invention, the thioredoxin and the disulfide bond isomerase are used as folding proteins, so that the tandem expressed mu-conopeptide can correctly and effectively form the mu-conopeptide with three pairs of disulfide bonds, and the method can effectively improve the yield of the mu-conopeptide. In addition, the [mu]-conopeptide fusion protein can be used for biosynthesizing [mu]-conopeptide, and compared with a traditional method, the [mu]-conopeptide fusion protein is more environmentally friendly, lower in cost compared with solid-phase synthesis and suitable for large-scale production.

This invention discloses a method for modifying the structure of recombinant mussel adhesive protein, comprising the following steps: S1. Optimizing the mussel adhesive protein gene sequence and cloning it into an expression vector to construct a recombinant plasmid; S2. Transforming the recombinant plasmid into competent cells and culturing them to obtain engineered bacteria; S3. Fermenting the engineered bacteria, collecting the bacterial cells by centrifugation, adding a lysis solution to break the bacterial cells, purifying and collecting the eluent, and dialysis to obtain a dialysate containing non-adhesive recombinant mussel adhesive protein; S4. Adding a chemical oxidant and an enhancer to the dialysate containing the non-adhesive recombinant mussel adhesive protein to react and obtain a highly adhesive recombinant mussel adhesive protein. This invention provides a novel chemical structure modification method for recombinant mussel adhesive protein. Through the synergistic effect of highly selective oxidants and enhancers, the tertiary structure of the protein is unfolded and the reaction activation energy is reduced, achieving efficient and simultaneous tyrosine hydroxylation and DOPA quinone crosslinking. The modification rate is more than 7 times higher than that of traditional chemical methods.

The present invention concerns Unfolded Protein Response (UPR) activating compounds and their use as a regenerative therapy for promoting thymic regeneration.