59 results about "Protein disulfide-isomerase" patented technology

The present disclosure describes methods and systems for improving the expression of a properly folded, biologically active protein of interest in a cell free synthesis system. The methods and systems use a bacterial cell free extract having an active oxidative phosphorylation system, and include an exogenous protein chaperone. The exogenous protein chaperone can be expressed by the bacteria used to prepare the cell free extract. The exogenous protein chaperone can be a protein disulfide isomerase and / or a peptidyl-prolyl cis-trans isomerase. The inventors discovered that the combination of a protein disulfide isomerase and a peptidyl-prolyl cis-trans isomerase produces a synergistic increase in the amount of properly folded, biologically active protein of interest.

The invention discloses a method for preparing oligomeric fucosylated glycosaminoglycan which is prepared by depolymerizing fucosylated glycosaminoglycan by a depolymerization method of peroxide catalyzed by a 4th period transition metal ion in an aqueous medium, and the preparation method has mild reaction condition, good reproducibility and stability, high pyrolysis selectivity and uniform and controllable product quality. The polysaccharide molecule number of the obtained oligomeric fucosylated glycosaminoglycan using GalNAc as a reducing end is not less than 80 percent, the weight averagemolecular weight is about 6, 000-20, 000Da, and the protein disulfide isomerase (PDI) is 1.0-2.0.

The present disclosure describes methods and systems for improving the expression of a properly folded, biologically active protein of interest in a cell free synthesis system. The methods and systems use a bacterial cell free extract having an active oxidative phosphorylation system, and include an exogenous protein chaperone. The exogenous protein chaperone can be expressed by the bacteria used to prepare the cell free extract. The exogenous protein chaperone can be a protein disulfide isomerase and / or a peptidyl-prolyl cis-trans isomerase. The inventors discovered that the combination of a protein disulfide isomerase and a peptidyl-prolyl cis-trans isomerase produces a synergistic increase in the amount of properly folded, biologically active protein of interest.

The invention provides compounds of formula (I) that inhibit PDI, for use in methods to treat or prevent a disease or condition in a subject that would benefit by inhibition of PDI. Formula (I)

The invention relates to a preparation method of a recombinant protein in the field of gene engineering and in particular relates to a preparation method of a recombinant human acidic fibroblast growth factor (haFGF) protein. The preparation method comprises the following steps: (1) designing primers according to the full-length sequence of the haFGF to obtain an original gene segment through cloning, and replacing a codon which exists in the original gene segment and is unfavourable to be expressed by escherichia coli, thus obtaining an optimized haFGF gene, wherein the nucleotide sequence of the optimized haFGF gene is shown in SEQ ID NO.1; cloning to obtain an original gene segment of a molecular chaperone PDI (protein disulfide isomerase), and replacing a codon which exists in the original gene segment and is unfavourable to be expressed by escherichia coli, thus obtaining an optimized PDI gene, wherein the nucleotide sequence of the optimized PDI gene is shown in SEQ ID NO.2; (2) linking and cloning the optimized haFGF gene, the optimized PDI gene, tag sequences and a protease cutting site sequence into a vector to obtain an efficient expression vector through construction; or firstly constructing standby vectors of part of the sequences, and then linking the remaining sequences on part of the sequences.

The present invention provides a method for treating or ameliorating the effects of a neurodegenerative disorder in a subject in need thereof. The method includes, for example, administering to the subject an effective amount of a compound selected from:combinations thereof, or an N-oxide, crystalline form, hydrate thereof, or a pharmaceutically acceptable salt thereof. The present invention also provides a method for treating or ameliorating the effects of a condition associated with increased protein disulfide isomerase (PDI) activity and a method of modulating PDI activity in a cell. The present invention also provides compounds, salts, compositions and kits useful for the provided methods.

The invention which belongs to the biotechnological field concretely relates to recombined hPDI491, a preparation method thereof and an application thereof. According to the invention, an RT-PCR process is applied to amplify to obtain hPDI491 cDNA, the hPDI491 cDNA is recombined with a Pichia pastoris expression vector pPICZalpha to obtain an expression plasmid pPICZalpha-hPDI, the pPICZalpha-hPDI is converted into Pichia pastoris X33, and high expression engineering strains are screened out. The expression of a large amount of the recombined hPDI491 in Pichia pastoris in secretion manner is realized. The purity of the engineering strains which are fermented, induction-expressed, and separation-purified is greater than 90%. Experiments proves that the hPDI491 has an in vitro correct folding promotion effect on other proteins.

The invention discloses a process method for synthesizing a lactic acid-lysine copolymer by catalytically opening a loop and copolymerizing with acetic bicyclo-guanidine (the molar content of diaminocaproic acid is 1-5 percent). The lactic acid-lysine copolymer is synthesized by taking acetic bicyclo-guanidine as a catalyst, taking lactide and lysine morpholine diketone as monomers, performing body loop opening copolymerization and undergoing a phenmethyl carboxide removing action. The process method has characteristics that: the acetic bicyclo-guanidine serving as a catalyst is bionic organic guanidinium which is effective, nontoxic and free from metals; the monomer transformation ratio is high (more than or equal to 95 percent); the yield is over 93 percent; a product does not contain any metal or other toxic residue, and has high biological safety; the number average molecular weight is adjustable in the range of 1.5-2.8*10<4>, and the molecular weight distribution is narrow (PDI (Protein Disulfide Isomerase) is less than or equal to 1.30); the molar content of lysine in the copolymer is adjustable in the range of 1-5 percent; and the synthesized lactic acid-lysine copolymer isan amphiphilic functional group biodegradable polymer, is suitable for serving as targeted and controlled release medicament carriers, and can be applied on other aspects in the field of biological medical tissue engineering.

The invention provides a pronuclei fusion expression vector which comprises a nucleotide fragment. The nucleotide fragment is composed of a promoter, a gene for coding a PDI (Protein Disulfide Isomerase) parasporal protein, and a joining region which are serially connected, wherein the joining region contains an enterokinase cleavage site and a multiple cloning site. The pronuclei fusion expression vector is capable of efficiently expressing an exogenous gene in a form of a fusion protein, is good in solubility of an expression product, simple in purification, and has natural bioactivity of the exogenous gene. The pronuclei fusion expression vector is suitable for mass production of a recombinant protein, is suitable for research and production of a genetic engineering product and a biological agent, and can also be used for researching a protein function and conformation relationship.

The invention discloses an expression system of fusion protein from human serum albumin and interleukin-1 receptor antagonist. The expression system comprises a host cell and an expression vector transferred into the same. The expression vector comprises a first expression vector with an inserted fusion protein gene and a second expression vector with an inserted protein disulfide isomerase gene.The fusion protein gene comprises the human serum albumin and the interleukin-1 receptor antagonist. The host cell is Pichia pastoris GS115. The co-expression host of PDI (protein disulfide isomerase) and IGH (immunoglobulin heavy) is established, and expression index of the IGH is evidently increased. The PDI is expressed intracellularly, the IGH secretory expression is subjected to extracellular secretory expression, and accordingly no newly generated other proteins occur when concentration of the IGH in collected medium supernate increases.

This invention relates to methods of expressing eukaryotic proteins in prokaryotic hosts, particularly eukaryotic proteins that require formation of disulfide bridges for biological activity. Various approaches are used including fusion to thioredoxin, cytoplasmic expression of disulfide isomerases, deficiencies in thioredoxin and / or glutathione reductases, deficiencies in proteases, and the like. The method is applicable to express monomeric and dimeric forms of the eukaryotic protein with biological activity such as monomeric and dimeric forms of a disintegrin or a disintegrin domain. Included are the vectors, host cells expressing the proteins, the expressed proteins and methods of using the proteins.