39 results about "Folding protein" patented technology

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.

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.

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.

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 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.

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 synthetic foldable protein has a tertiary structure emulating the tertiary structure of a reference foldable protein. The reference foldable protein has a folding nucleus peptide sequence associated with folding the reference foldable protein. The synthetic foldable protein also has a peptide sequence including the folding nucleus peptide sequence and at least one repeat thereof.

The invention provides an expression cassette of lipoprotein lipase, a recombinant expression vector, a recombinant strain and application of the recombinant strain in production of lipoprotein lipase, and belongs to the technical field of gene engineering. The invention provides an expression cassette of lipoprotein lipase. The expression cassette comprises the following gene elements which are connected in series: an arabinose inducible promoter, double-copy lipoprotein lipase lipA and single-copy folded protein lipB. According to the invention, an expression element optimization strategy is adopted, an inducible promoter is adopted to replace a constitutive promoter to accurately regulate and control expression, and a specific site double-copy integration strategy and co-expression are combined to participate in protein folding genes, so that efficient secretory expression of lipoprotein lipase in host cells is realized.

This invention provides the application of a combination of biomarkers for early screening and diagnosis of transthyretin-mediated cardiac amyloidosis (ATTR-CA) in the preparation of products for early screening and diagnosis of ATTR-CA in high-risk populations. Generally, one or more biomarkers are used, such as total prealbumin (total PA), transthyretin (TTR) tetramer, misfolded proteins, protein misfolding rate, and soluble growth-stimulating gene 2 protein (sST2). Significant differences in these biomarkers are observed between high-risk and non-high-risk ATTR-CA populations, suggesting an association between the biomarkers and ATTR-CA. The combined use of multiple biomarkers demonstrates high sensitivity and specificity in early screening and diagnosis of ATTR-CA in high-risk populations, exhibiting high diagnostic efficacy and providing new ideas and strategies for the early diagnosis and treatment of ATTR-CA.

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 method for diagnosing or predicting preeclampsia in a pregnant woman or determining whether a pregnant woman is at risk of developing preeclampsia is provided, comprising: a) obtaining at least one (such as two or more) maternal or fetal parameter or factor of the pregnant woman selected from the group comprising: (i) one or more of mean arterial pressure, BMI at sampling, ΔBMI, gestational age, and ΔBMI / GA; (ii) maternal age; (iii) one or more of urinary protein level and ratio of urinary protein level (such as total urinary protein) to urinary creatinine level; (iv) one or more of history of preeclampsia in previous pregnancies and having received antihypertensive medication; and (v) any combination of (i)-(iv), b) determining whether the pregnant woman has misfolded proteins or misfolded protein aggregates in her urine, and c) combining the at least one maternal or fetal parameter or factor obtained in step a) with the level of misfolded proteins or misfolded protein aggregates in the pregnant woman's urine to diagnose or predict preeclampsia or determine the risk of preeclampsia.

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 invention discloses a method for screening a bacterial membrane protein folding machine targeting inhibitor based on fluorescence complementation, and belongs to the field of biological medicine detection and analysis. The invention provides a report vesicle system, which comprises a report vesicle and a protein inclusion body, the report vesicle is a vesicle in which LgBiT and luciferase luminescent substrates are stored and an outer membrane is provided with a BamABCDE complex; the protein inclusion body is a to-be-folded protein connected with a HiBiT tag in front of a passenger structure. According to the report vesicle system, the folding process of the outer membrane protein of bacteria can be monitored in a vesicle simulated lipid membrane environment in a real-time, sensitive and high-throughput manner, and on the basis, a method and a system for rapidly and effectively screening a targeted BAMABCDE complex inhibitor are provided.