338results about "Fusions for enhanced expression stability/folding" patented technology

The invention belongs to the field of biological medicines, and particularly relates to a varicella-zoster virus gE fusion protein as well as a preparation method and application thereof. The varicella-zoster virus gE fusion protein comprises an amino acid sequence as shown in SEQ ID NO: 1 and at least one of the following fragments: a signal peptide, a Foldon sequence, a Ferritin sequence and a His tag sequence, the signal peptide can improve the protein expression quantity, the Foldon sequence can improve the stability of the protein, the particle size of an antigen can be increased, the immunogenicity of the antigen can be improved, and the fusion protein can be used for preparing a medicine for treating the varicella-zoster virus gE fusion protein. The Ferritin sequence can enhance immune response, a vaccine prepared from the gE fusion protein has excellent immunogenicity, and compared with a commercially available vaccine Shingrix, higher humoral immunity and cellular immunity levels can be induced.

The invention belongs to the field of synthetic biology, and particularly relates to collagen fusion polypeptide as well as application and a preparation method thereof. The collagen fusion polypeptide provided by the invention comprises: (a) a first polypeptide of recombinant III-type collagen; and (b) a second polypeptide comprising a trimerized folding domain. The collagen fusion polypeptide provided by the invention has the advantages of low immunogenicity, good biocompatibility, high biological activity and the like, can significantly promote proliferation and migration of human skin fibroblasts, and has stronger ability of promoting regeneration of endogenous collagen. The collagen fusion polypeptide has a triple helix structure similar to natural collagen, and can be self-assembled to form collagenous fibers which are good in morphology and similar to natural collagen. The method can be widely applied to the fields of skin repair dressings, biological materials, artificial implants, medical instruments, cosmetics, health care products, food and the like.

The invention relates to the technical field of gene engineering, in particular to Escherichia coli for efficient soluble expression of recombinant human collagen III and construction and application thereof. The invention constructs Escherichia coli for efficient soluble expression of recombinant human III type collagen, and the Escherichia coli can co-express the recombinant human III type collagen and molecular chaperone which are regulated and expressed by signal peptide. Experiments prove that the engineering bacterium provided by the invention has the capability of efficient soluble expression of the recombinant human III-type collagen, and the recombinant human III-type collagen provided by the invention has good stability and anti-inflammatory effect.

The present invention provides compositions and methods comprising an activator of interleukin-18 (IL-18) activity for use in therapeutic and non-therapeutic applications. The activator provides IL-18 signaling activity even in the presence of an inhibitory molecule 5 such as IL-18 binding protein (IL-18BP).

Provided herein are compositions and methods for adoptive cell therapy comprising engineered immune cells that express a tumor antigen-targeted chimeric antigen receptor and a prodrug converting enzyme.

Provided are a recombinant human interleukin-11 mutant, and a conjugate of the mutant conjugated to a chemical molecule and a use thereof. The recombinant human interleukin-11 mutant achieves soluble expression in an Escherichia coli expression system by means of an SUMO sequence tandem fusion method. The conjugate is obtained by the site-specific conjugation of the mutant to the chemical molecule at a mutant amino acid residue in position W147 by means of a covalent bond. The chemical molecule is at least one of polyethylene glycol, an alkanoic acid aliphatic chain, and a hydrophilic high-molecular polymer containing an alkanoic acid aliphatic chain. The conjugate retains the binding capacity to human interleukin-11 receptor A while demonstrating significantly reduced binding capacity to human IL-6 signal transducer receptor. The conjugate shows the ability to block the signaling between human IL-11 and IL-11 RA and between human IL-11 and GP130 either in vivo or in vitro and can be used for the treatment of associated diseases with tissue fibrosis as a clinical manifestation caused by elevated primary or secondary IL-11.

The present disclosure provides engineered alpha klotho polypeptides and methods of their production and use. Soluble alpha klotho polypeptides disclosed herein comprise an alpha klotho moiety having one or more mutations for enhancing activity, production yield, and / or stability relative to human alpha klotho. The disclosure further provides pharmaceutical compositions comprising the engineered alpha klotho polypeptides, and methods of use of the engineered alpha klotho polypeptides in treatment methods, including treatment of age- related conditions and kidney disease. Also disclosed are nucleic acids encoding the engineered alpha klotho polypeptides, recombinant cells that express the engineered alpha klotho polypeptides, and methods of producing the engineered alpha klotho polypeptides.

The invention discloses a method for improving the solubility and thermal stability of sweet protein, and belongs to the technical field of biosynthesis, the method comprises the following steps: constructing a gene tandem recombinant plasmid containing a monellin x-3C-sfGFP-3C-monellin y expression cassette, x is greater than or equal to 1, y is greater than or equal to 1, 5 is greater than or equal to x + y is greater than or equal to 3, a 3C protease recognition sequence is also inserted between adjacent copies of monellin, and each monellin is connected with a purification tag; the gene tandem recombinant plasmid is transformed into escherichia coli and inducible expression is carried out, thalli are split and purified to obtain fusion protein, 3C protease is used for enzyme digestion, and the target protein is obtained after re-purification. The total protein yield and the solubility proportion are remarkably improved by adopting a series construction mode of plasmids, meanwhile, the stability of monellin is effectively improved, the purification step is simpler and more convenient, and the method is suitable for industrial production.

The invention belongs to the technical field of genetic engineering, and particularly relates to a mutant P17 oligopeptide and application thereof in preparation of a nano antibody. The amino acid sequence of the mutant P17 oligopeptide is KGFRDEKKRFKNTKG. The mutant P17 oligopeptide is a mutant P17 oligopeptide. The mutant P17 oligopeptide provided by the invention has good chaperone-like activity and dissolution promoting effect, has good applicability to different nano antibody strains, is fused with a nano antibody and is expressed in a prokaryotic expression system, so that the solubility and expression efficiency of the nano antibody in the prokaryotic expression system can be effectively improved, and the application prospect is broad. The soluble yield of the nano antibody is greatly improved; moreover, the mutant P17 oligopeptide is beneficial to keeping the biological activity of the antibody, the nano antibody with high activity and high expression quantity is obtained, favorable conditions are created for large-scale production of the nano antibody, and the mutant P17 oligopeptide has a wide application prospect.

The invention discloses recombinant yarrowia lipolytica with high yield of 3-fucosyllactose as well as a construction method and application of the recombinant yarrowia lipolytica. According to the invention, a reinforced lactose transporter lac12 gene, a reinforced GDP-mannose-4, 6-dehydratase gmd gene, a reinforced GDP-L-fucose synthase gmer gene and a 3-fucosyltransferase SUMO-fut3Bc gene with a reinforced SUMO tag are subjected to integrated expression on a recombinant yarrowia lipolytica chromosome, and a 6-phosphogluconolactonase pgl gene is subjected to overexpression, so that a recombinant yarrowia lipolytica strain is obtained. The constructed recombinant yarrowia lipolytica can maintain the genetic stability of a target gene, the yield and production efficiency of 3-fucosyllactose are remarkably improved, and the requirements of industrial production can be met.

The invention discloses a method for improving soluble expression of non-ribosome peptide synthetase in escherichia coli. According to the method, a polyion oligopeptide label is connected to the N end or the C end of the non-ribosome peptide synthetase; the method comprises the following steps: adding a polyion oligopeptide tag (D) 5 at the N / C end of ebony in a fusion protein SGE formed by non-ribosomal peptide synthetase ebony for producing decarboxylated carnosine and phosphokinase Sfp through a connecting peptide GSG, or replacing a GSG sequence in an original SGE sequence with the polyion oligopeptide tag (D) 5 to construct new plasmids, and transforming the obtained three new plasmids in recombinant escherichia coli to obtain the decarboxylated carnosine-producing non-ribosomal peptide synthetase ebony-phosphokinase Sfp. The solubility of the non-ribosome peptide synthetase in escherichia coli is greatly improved, and the addition of the polyion oligopeptide tag (D) 5 at the N end is more beneficial to reducing the formation of inclusion bodies and improving the yield of decarboxylated carnosine.

The invention relates to a recombinant III-type human collagen expression strain with a whole chain length and a triple helix structure as well as a preparation method and application thereof, and belongs to the technical field of genetic engineering bacteria. The recombinant pichia pastoris genetically engineered bacterium expresses recombinant III-type human collagen with a full chain length and a triple helix structure, and a ku70 gene of the recombinant pichia pastoris genetically engineered bacterium is knocked out and knocked in a recombinant 4-proline hydroxylase coding gene and an FKBP22 coding gene. The recombinant pichia pastoris genetically engineered bacterium provided by the invention can be used for obtaining the recombinant full-chain-length triple-helix recombinant III-type human collagen with a higher expression level through high-density fermentation, does not contain heterologous sequences, has higher hydroxyproline content and thermal stability, is close to natural human collagen, has a stable triple-helix structure, is high in thermal stability, and can be used for preparing the recombinant III-type human collagen. The application in the fields of medical health and medical beauty such as medical beauty filling and tissue repair can be met.

The invention discloses a method for improving the heterologous soluble expression efficiency of cytochrome P450, which is characterized in that plasmids containing coding genes of fungus-derived cytochrome P450 truncated protein and plasmids containing molecular chaperones are jointly transferred into host bacteria to obtain recombinant genetically engineered bacteria for improving the heterologous soluble expression efficiency of cytochrome P450. Through host screening, molecular chaperonin and cytochrome P450 gene modification, the soluble expression efficiency of cytochrome P450 in Escherichia coli is improved, and the problems of poor protein solubility and low expression efficiency when CYP450 is expressed in Escherichia coli at present are solved.

The invention discloses an immunoregulatory factor composition, which comprises a first component and a second component, the first component is a type I cytokine or a long-acting form thereof, and the second component is a type II cytokine or a long-acting form thereof; the long-acting form is formed by fusing a cell factor with a fusion partner through a connecting peptide or performing PEGylation modification on the cell factor, and the fusion partner is an Fc fragment of IgG or human serum albumin or an engineered mutant of one of the Fc fragment and the human serum albumin. The immunomodulatory factor composition synergistically activates precursor depleted CD8 + T cells and terminal depleted CD8 + T cells through a combination therapy of type I and type II cytokines. The invention also discloses nucleic acid, a carrier and a cell related to the immunomodulatory factor composition. The invention also discloses an application of the immunoregulatory factor composition in preparation of drugs for treating cancers. The immunoregulatory factor composition can relieve the depletion state of depleted T cells and improve the effector function and proliferation capacity of the depleted T cells so as to control tumor progression.

Described herein are engineered paraneoplastic Ma protein (PNMA) capable of forming a capsid. In some embodiments, the engineered PNMA proteins comprise one or more modifications that enhance binding or loading of a cargo into the capsid, one or more modifications that modify cell-specificity of the capsid, one or more modifications that enhance intracellular delivery of the capsid, or a combination thereof. Also described herein are delivery systems comprising capsids comprising an engineered PNMA protein and a cargo.

Provided are a GLP-1GIP analog, an FGF21 protein, a trifunctional protein, and use thereof. The trifunctional protein comprises the GLP-1 / GIP analog and the FGF21 protein. Further provided is use of the GLP-1 / GIP analog, the FGF21 protein, and the trifunctional protein in the preparation of a drug for preventing and / or treating diseases related to GLP-1, GIP, and / or FGF21. The provided GLP-1 / GIP analog, FGF21 protein, and trifunctional protein can synergistically regulate blood glucose and lipid levels in vivo, meet multiple needs in patients with type 2 diabetes, including lowering blood glucose, alleviating hepatic steatosis, promoting weight loss, ameliorating circulating lipid metabolism disorder, etc., and have better biological safety and tolerability and longer in vivo half-lives.

The present invention relates to a fusion protein comprising a main protein and one or more extension peptides, wherein the amino acid sequence of the main protein is identical or similar to the amino acid sequence of a mammalian protein or a fragment thereof, and the extension peptides comprise a cluster of O-glycosylated amino acids. The extension peptide is identical to a non-repeated sequence of a mammalian protein and / or is identical or similar to SEQ ID NO:1. The main protein is preferably VWF. The fusion protein has an increased half-life compared to the main protein and can be used to increase the half-life of a binding partner, such as FVIII. The invention further relates to complexes formed by the fusion proteins, polynucleotides encoding the fusion proteins, and vectors and host cells containing the polynucleotides. [Selected Figure] Figure 1

Provided are a method for constructing a soluble expression plasmid mutant library, and the use thereof. The method for constructing the soluble expression plasmid mutant library comprises: introducing mutations into a ribosome binding site on a soluble expression plasmid, a sequence composition and length of a region between the ribosome binding site and an initiation codon of a fusion protein to be expressed, and a translation initiation region, so as to obtain a mutant library. By means of the method, plasmid mutants with increased expression levels are obtained, thereby increasing the expression level of a target fusion protein and the final yield of a target polypeptide. The present invention solves the problem of low soluble expression levels of certain polypeptides in Escherichia coli.

The disclosed method of treating a malignant solid tumor in a subject includes the steps of administering to the subject an immunomodulatory dose of a radioactive phospholipid ether metal chelate, a radiohalogenated phospholipid ether, or other targeted radiotherapy (TRT) agent that is differentially retained within malignant solid tumor tissue, and either (a) performing in situ tumor vaccination in the subject by introducing into at least one of the malignant solid tumors one or more agents capable of stimulating specific immune cells within the tumor microenvironment, or (b) performing immunotherapy in the subject by systemically administering to the subject an immunostimulatory agent, such as an immune checkpoint inhibitor. In a non-limiting example, the radioactive phospholipid ether metal chelate or radiohalogenated phospholipid ether has the formula:wherein R1 comprises a chelating agent that is chelated to a metal atom, wherein the metal atom is an alpha, beta or Auger emitting metal isotope with a half-life of greater than 6 hours and less than 30 days, or wherein R1 comprises a radioactive halogen isotope. In one such embodiment, a is 1, n is 18, m is 0, b is 1, and R2 is —N+(CH3)3.

The invention relates to the technical field of biology, in particular to a mirror image coupled protein nanoparticle conjugate and application thereof. The conjugate comprises: ii) a functional protein labeled by a first peptide tag, and ii) a scaffold protein labeled by a second peptide tag, the first peptide tag and the second peptide tag being composed of D-amino acid residues; the first peptide tag and the second peptide tag form a peptide partner pair, and the first peptide tag and the second peptide tag are coupled and combined through an isopeptide bond. The conjugate provided by the invention is easier to escape from the recognition of protease, can obviously prolong the half-life period in a biological system, is not easy to cause unnecessary immunoreaction, and has good biological stability and safety.

The present invention relates to research tools for structural biology, particularly for facilitating the determination of three-dimensional structures of biological macromolecules. More specifically, the present invention helps to improve the overall feasibility of structure determination, resulting in higher resolution and better quality of three-dimensional structures of proteins through complex formation with novel fusion polypeptides.

It is an object of the present invention to provide a method of reliably chemogenetically controlling the activity of a protein, and to provide a protein used in the present method. Specifically, the present invention relates to a plurality of protein fragments, which recover the activity of the protein when the plurality of protein fragments assemble, and in which each of the plurality of protein fragments is tagged with a degradation domain (DD) sequence. In addition, the present invention also relates to a method of regulating the activity of a protein in a cell, comprising: a step of introducing into a cell, a fusion of a DD sequence, and each fragment of a protein that has been split into fragments, such that the fragments recover the activity of the protein when they assemble; and a step of introducing a DD sequence-specific stabilizing factor into the cell.

The present disclosure relates generally to recombinant polynucleotides comprising novel pro-region DNA sequences. Certain aspects of the disclosure relate to recombinant Gram-positive bacterial strains comprising one or more introduced polynucleotides comprising novel pro-region DNA sequences operably linked to a DNA sequence encoding a protein of interest.

The present invention relates to antigens, more particularly protein antigens of enterococcal pathogens derived from the protein AdcA that are particularly useful as vaccine components and constructs for therapy and / or prophylaxis of bacterial infections. The present invention further relates to antibodies directed against the antigens of the invention and medical uses thereof.

This invention discloses a functional peptide for increasing recombinant protein yield, the sequence of which is shown in any of SEQ ID No. 1-15. The encoding gene and its applications are also disclosed, along with corresponding methods for increasing protein yield. The functional peptide of this invention can significantly increase the yield of recombinant proteins, significantly reduce the loss rate of recombinant proteins during expression and purification in the MtuΔI-CM protein expression system, reduce the production cost of recombinant proteins, and improve the stability of the target protein. It is suitable for the production of various recombinant human proteins, laying the foundation for large-scale, low-cost mass production of various recombinant proteins and possessing good commercial application prospects.

A polypeptide tag is provided, having the amino acid sequence Xaa1Xaa2Xaa3PHDYNXaa4Xaa5Xaa6, where Xaa1, Xaa2, Xaa3, Xaa4, Xaa5, and Xaa6 are each independently an amino acid or absent, and the polypeptide tag is used to label a target protein. A second aspect provides a polypeptide fusion protein, comprising two structures: (1) any of the polypeptide tags of the first aspect and (2) a target protein linked to the polypeptide tag. A cell-free protein synthesis system and its use in in vitro protein synthesis are also provided. By constructing a fusion protein from a polypeptide tag and a target protein, the expression level of the labeled target protein can be effectively increased even if the polypeptide tag is not removed.

The present invention provides a method of modulating a molecular mass of a fusion polypeptide, wherein the fusion polypeptide comprises a target polypeptide fused to a tag suitable for affinity purification or detection of the target polypeptide, wherein the tag comprises at least one constant domain of an immunoglobulin light chain or orthologous polypeptide or fragment thereof and wherein the molecular mass and / or an isoelectric point of the fusion polypeptide is modulated by increasing the molecular mass of the tag via combining the one constant domain of an immunoglobulin light chain or orthologous polypeptide or fragment thereof with one or more further constant domain(s) of an immunoglobulin light chain or orthologous polypeptide(s) or fragment(s) thereof; wherein the constant domain of the immunoglobulin light chain or orthologous polypeptide or fragment thereof of the tag is monomeric; and wherein optionally the target polypeptide is fused to the tag via a first linker. The invention further provides a fusion polypeptide obtained or obtainable by this method. The invention also provides a tag and a fusion polypeptide comprising the tag.

The present invention belongs to the field of biomedicine, and specifically relates to a recombinant subunit vaccine against respiratory syncytial virus (RSV) and its application, and more specifically to a recombinant fusion protein comprising the Head only (RHF) domain of RSV envelope fusion protein F and a multimerization domain such as an immunoglobulin Fc fragment, an expression construct comprising the recombinant fusion protein, a method for preparing the recombinant fusion protein, and an immunogenic composition comprising the recombinant fusion protein, such as a vaccine.

Provided herein are compositions and methods for adoptive cell therapy comprising engineered immune cells that express a tumor antigen-targeted chimeric antigen receptor and a prodrug converting enzyme.

Disclosed are compositions and methods for treating a disease or disorder such as cancer in a subject in need thereof. In some embodiments, the methods include administering to the subject a vector that has a first nucleic acid sequence encoding a promoter operably linked to each of a second nucleic acid sequence encoding a therapeutic polypeptide, and a third nucleic acid sequence encoding a peptide domain that is stabilized when phosphorylated by kinase activity in a target cell and / or tissue. In some embodiments, the target cell and / or tissue can be a cell and / or tissue undergoing a stress response. In some embodiments, the target cell and / or tissue can be a cell and / or tissue in which a CK2 kinase is active. The kinase activity can be elevated extracellular regulated kinase (ERK) activity, p38 MAP kinase activity, and / or CK2 activity.