18 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 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.

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

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.

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.

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.

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.

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.

The invention discloses an engineered OST1 signal peptide which is a mutant formed by replacing or inserting residues VSS at 19-21 sites in a junction region of an H region and a C region of a wild type OST1 signal peptide with an amino acid sequence shown as SEQ ID NO: 1 by acidic residues D and E, and a composite secretion signal peptide formed by the engineered OST1 signal peptide and an alpha-factor Pro section connected to the C terminal of the engineered OST1 signal peptide, the method is used for driving secretory expression of foreign proteins in yeast, can significantly improve the secretory efficiency of high-electric-point proteins, glycoproteins and complex folded proteins, and has popularization and application prospects.

The invention provides a process for manufacturing certolizumab pegol from bacterial host cells. The invention provides refolding process of certolizumab wherein the solubilized solution of heavy chain and light chain of certolizumab is treated with refolding buffer under suitable conditions including pII, temperature, and incubation period to obtain high quality and quantity of refolded protein. The invention further provides efficient pegylation process to obtain certolizumab pegol in good yields.

The application discloses a method for improving expression of exogenous glycoprotein by yeast, comprising the following steps: without changing endogenous genes of a yeast host cell and without introducing exogenous sugar chain processing enzymes, an exogenous N-acetylglucosamine transferase I (GnT1) gene from different species is integrated into a genome of a Pichia pastoris host bacterium for expressing glycoprotein, and a Pichia pastoris engineering bacterium for expressing the exogenous GnT1 is constructed, so that the expression amount of the exogenous glycoprotein in the Pichia pastoris engineering bacterium is significantly improved, the proportion of high mannose type N-glycan is reduced, and the proportion of complex type N-glycan is increased. The method is suitable for biosynthesis of polysaccharide glycosylation protein, high isoelectric point protein and complex folding protein, and has good popularization and application prospect in the fields of biological pharmacy, functional protein, nutritional and healthy raw material and industrial biological manufacturing.

Methods and kits are provided for amplifying and detecting αS proteins from samples, for example, from patients having Parkinson's Disease. For example, a method for determining a presence of a soluble, misfolded αS protein may include: contacting the sample with a monomeric, folded αS protein to form an incubation mixture; conducting an incubation cycle two or more times on the incubation mixture effective to form an amplified portion of misfolded αS protein; incubating the incubation mixture effective to cause misfolding and / or aggregation of at least a portion of the monomeric, folded αS protein in the presence of the soluble, misfolded αS protein; physically disrupting the incubation mixture effective to at least partly de-aggregate at least a portion of a misfolded αS aggregate present; and determining the presence of the soluble, misfolded αS protein in the sample by detecting at least a portion of the soluble, misfolded αS protein.

The invention provides a photoacoustic enhanced microcavity misfolded protein detection system and method.The photoacoustic enhanced microcavity misfolded protein detection system comprises a micro laser cavity unit, the micro laser cavity unit is a liquid crystal micro laser cavity doped with a quantum dot material with the fluorescence lifetime larger than or equal to 10 ns, and the liquid crystal micro laser cavity takes a cationic surface active agent solution as a water phase; a liquid crystal material is mixed with a quantum dot material to serve as an oil phase, the surface of the oil phase is modified with an aptamer for specifically capturing misfolded protein, and the oil phase is used for generating WGM resonance light; a pulse laser excitation unit; the photoacoustic material unit is used for responding to photoacoustic excitation to generate photoinduced ultrasonic waves; the photoacoustic excitation unit is used for exciting the photoacoustic material unit, generating controlled photoinduced ultrasonic waves, acting on the liquid crystal micro laser cavity and generating a modulation effect on WGM resonant wavelength drift caused by misfolded protein binding; and the spectrum detection unit is used for collecting a spectrum signal of the WGM resonance light drifted and modulated by the photoacoustic excitation unit.