171 results about "C-terminus" patented technology

The invention discloses a recombinant humanized XVII type collagen as well as an expression vector and a genetically engineered bacterium thereof. Four functional fragments with high activity are obtained through screening, one of the fragments is optimized and then combined according to a certain sequence to obtain recombinant humanized XVII type collagen JYC1703, or one of the fragments is optimized and then combined according to a certain sequence, and GPP sequences are fused at an amino terminal and a carboxyl terminal respectively to obtain recombinant humanized XVII type collagen JYC1701 and JYC1702, and the recombinant humanized XVII type collagen JYC1701 and JYC1702 are combined according to a certain sequence to obtain the recombinant humanized XVII type collagen JYC1701 and JYC1702. After codon optimization, amplifying and cloning into an expression vector, transforming into pichia pastoris competent cells after enzyme digestion linearization, and screening high-copy recombinants through G418 resistance gradient, so as to obtain pichia pastoris genetically engineered bacteria for expressing recombinant humanized XVII type collagen. The recombinant humanized XVII type collagen JYC1701, JYC1702 and JYC1703 disclosed by the invention show excellent functions of promoting cell adhesion, cell proliferation and cell migration, and are suitable for preparing products such as skin care products or medical instruments and the like.

Compositions and methods relating to paraoxonase fusion polypeptides are disclosed. In some aspects, the fusions are bispecific molecules that include a first biologically active polypeptide linked amino-terminal to a biologically active paraoxonase, wherein the first biologically active polypeptide is a DNase, an RNase, a SOD1, a CTLA-4 extracellular domain, a CD40 extracellular domain, or a polypeptide that specifically binds and neutralizes an inflammatory cytokine. Bispecific fusions may further include a second biologically active polypeptide (e.g., a dimerizing or FcRn-binding domain) linked carboxyl-terminal to the first biologically active polypeptide and amino-terminal to the paraoxonase. In other aspects, a fusion polypeptide includes a biologically active paraoxonase linked carboxyl-terminal or amino-terminal to a dimerizing or FcRn-binding domain. Also disclosed are dimeric proteins comprising first and second paraoxonase fusion polypeptides as disclosed herein. The fusion polypeptides and dimeric proteins are useful in methods for therapy.

The invention belongs to the technical field of gene engineering, and particularly relates to a strain for efficiently expressing beta-lactoglobulin as well as a construction method and application of the strain. By utilizing the beta-lactoglobulin expression vector disclosed by the invention, beta-lactoglobulin is guided to be secreted out of a host cell by an exogenous signal peptide, and efficient secretory expression of the beta-lactoglobulin in the host cell is realized by integrating a multi-copy expression cassette through a genome and co-expressing ATP enzyme Ssa4 participating in protein folding and coping with pressure; the amino terminal and the carboxyl terminal of the secreted and expressed beta-lactoglobulin do not contain redundant amino acids, and do not contain animal-derived components. Meanwhile, the invention provides a construction method and a fermentation culture method of the engineering strain, compared with a single-copy genetically engineered strain, the expression quantity of the genetically engineered strain in shake-flask fermentation is increased by 2.47 times, and the genetically engineered strain has a good industrial application prospect.

The invention belongs to the technical field of biology, and particularly relates to antibacterial peptide based on large yellow croaker CXCL8 protein and application of the antibacterial peptide. The preparation method comprises the following steps: by taking a large yellow croaker CXCL8 protein as a template, intercepting a linear polypeptide of 13 amino acids, and carrying out carboxyl terminal amidation modification, so as to obtain the antibacterial peptide AVV13N, and the amino acid sequence of the antibacterial peptide AVV13N is Ala-Arg-Trp-Val-Arg-Arg-Val-Leu-Lys-Lys-Met-Val-NH2; the antibacterial peptide AVK13N is obtained by interchanging lysine at the 11th site and valine at the 13th site in an amino acid sequence of the antibacterial peptide AVV13N, and the amino acid sequence of the antibacterial peptide AVK13N is Ala-Arg-Trp-Val-Arg-Arg-Val-Leu-Lys-Lys-Val-Met-Lys-NH2. The molecular weights of the antibacterial peptides AVV13N and AVK13N are both 1669.12 Da, the isoelectric points of the antibacterial peptides AVV13N and AVK13N are both 12.44, and the antibacterial peptides AVV13N and AVK13N are both alkaline polypeptides. The antibacterial peptides AVV13N and AVK13N have broad-spectrum bactericidal activity and can be used for specifically killing gram-positive bacteria and gram-negative bacteria. In addition, the antibacterial peptides AVV13N and AVK13N have relatively weak hemolytic activity, relatively low cytotoxicity and good temperature stability, pH value stability and salt ion stability. The antibacterial peptides AVV13N and AVK13N provided by the invention can replace antibiotics and are applied to prevention and treatment of bacterial diseases.

The invention relates to a second antibody for resisting mouse IgG and rabbit IgG as well as a preparation method and application of the second antibody. The second antibody of the anti-mouse IgG and the rabbit IgG sequentially comprises an anti-mouse IgG nano antibody, a biotin fixed-point binding peptide fragment and an anti-rabbit IgG nano antibody from an amino terminal to a carboxyl terminal. According to the design of the second antibody of the anti-mouse IgG and rabbit IgG with a brand-new structure and the preparation method, high-efficiency expression is realized, the second antibody of the anti-mouse IgG and rabbit IgG capable of realizing fixed-point biotinylation is obtained, the second antibody can be further subjected to various modification reformation based on a biotin-streptavidin bridge, the preparation method of the second antibody of the polymerase labeled anti-mouse IgG and rabbit IgG is further developed, and the application of the second antibody of the polymerase labeled anti-mouse IgG and rabbit IgG is expanded. According to the method, the polymerase-labeled mouse-rabbit universal secondary antibody with similar affinity to mouse IgG and rabbit IgG is obtained, the preparation period of the secondary antibody can be shortened, the preparation process of the secondary antibody is simplified, the stability between batches is improved, and meanwhile, the dependence on experimental animals is reduced.

The invention belongs to the technical field of polypeptide drugs in biochemistry, and discloses three antibacterial peptide analogues as well as a preparation method and application thereof, the sequence of an antibacterial peptide P-1 is CH3CO-GGGENIKKILSKIKLLK-NH2, the sequence of an antibacterial peptide P-2 is CH3CH2CH2CO-GGGENIKKILSKIKLLK-NH2, the sequence of an antibacterial peptide P-3 is CH3 (CH2) 6CO-GGGENIKKILSKIKLLK-NH2, the carboxyl ends of polypeptides are subjected to amidation, and the polypeptides are all positively charged when the pH is equal to 7. The artificially synthesized antibacterial peptide disclosed by the invention shows a remarkable sterilization effect on multidrug resistant staphylococcus epidermidis. P-1, P-2 and P-3 have small molecular weight, are convenient to synthesize, have broad-spectrum bactericidal activity, are self-assembled into nanoparticles, have low cytotoxicity, resist trypsin degradation and the like, and have wide application prospects.

Bacteriophages that comprise a tail fiber protein sequence comprising a C terminus host recognition site described by SEQ ID NO: 1 and which infect Ralstonia solanacearum which causes bacterial wilt disease are disclosed. Compositions comprising these bacteriophages may be employed as wilt control agents. Methods for preventing or treating diseases caused by Ralstonia solanacearum such as bacterial wilt disease are also disclosed. A bacterial wilt disease control method in a plant may include administering such bacteriophages to a plant or to a plant growth medium and can be effective against a wide variety of Ralstonia solanacearum strains. Nucleic acids encoding the C terminus host recognition site and amino acid sequences comprising the host recognition site are also disclosed.

Disclosed herein are methods for treating or reducing symptoms of a neurodegenerative disease in a subject. This disclosure relates to a method of restoring the tertiary structure of a mutant C terminus of HSP70-Interacting Protein (CHIP) to the tertiary structure of wild-type CHIP. Also disclosed herein are methods of increasing ubiquitination of a protein by a mutant C terminus of HSP70-Interacting Protein (CHIP) and methods of treating a neurodegenerative disorder comprising administering a therapeutically effective amount of a small molecule or a peptide to a subject.

The invention discloses an IL10-secreting and BAFFR-targeting chimeric antigen receptor, a CAR-T cell and application. The IL10-secreting and BAFFR-targeting chimeric antigen receptor comprises a signal peptide region, a BAFFR-targeting antigen binding domain, a hinge region, a transmembrane structural domain, a co-stimulation structural domain, a signal transduction structural domain and human IL10 which are sequentially connected from an amino terminal to a carboxyl terminal, wherein the signal peptide region, the BAFFR-targeting antigen binding domain, the hinge region, the transmembrane structural domain, the co-stimulation structural domain, the signal transduction structural domain and the human IL10 are sequentially connected from the amino terminal to the carboxyl terminal; the co-stimulation structural domain comprises a CD28 co-stimulation structural domain and a 4-1BB co-stimulation structural domain which are connected, and the signal transduction structural domain is ITAM1 of CD3 zeta. The kit can specifically aim at autoimmune B cells, and immunodeficiency and patient infection caused by full B cell deletion are avoided. The secretion of IL10 on the CAR-T cells can reduce the depletion rate of the CAR-T cells in vivo, so that the long-acting inhibition effect on the disease process is achieved.

Provided is a polypeptide which contains a polypeptide having at least 80%, 85%, 90%, 95%, or 99% identity to amino acid sequence MTYRPGYNPFG (SEQ ID NO: 41) or amino acid sequence AKGYYRPYGVPV (SEQ ID NO: 22), or a polypeptide having one or more amino acid substitutions, deletions and / or additions relative to the amino acid sequence as shown in SEQ ID NO: 41 or 22. The provided polypeptide can interfere with the binding of a Brag2 synaptic protein to the C terminus of an AMPA receptor, thereby inhibiting NMDA-mediated excessive endocytosis of an AMPA receptor and neuronal apoptosis, and playing a role in protecting neuronal cell activity. Therefore, the polypeptide has application prospects and potential development values as a drug for treating nervous system diseases such as strokes, depression, Alzheimer's disease and drug addiction.

The invention discloses mucosal immune-enhanced recombinant lactobacillus for expressing PEDV (porcine epidemic diarrhea virus) S1 protein as well as a construction method and application of the mucosal immune-enhanced recombinant lactobacillus. The mucosal immune-enhanced recombinant lactobacillus contains a recombinant lactobacillus expression vector for expressing PEDV S1 protein, the amino acid of the PEDV S1 protein is fused with M cell targeting peptide (Co1) and dendritic cell targeting peptide (6aa), the carboxyl terminal of the PEDV S1 protein is fused with a mucosal immune adjuvant (LTB), and the connection sequence of all the parts is Co1-6aa-S1-LTB. According to the mucosal immune enhanced recombinant lactobacillus constructed by the invention, through the synergistic effect of the targeting peptide (DC / MC targeting peptide) and the adjuvant (LTB), the mucosal immune response efficiency (including intestinal mucus SIgA and serum IgG levels) of a PEDV S1 antigen is remarkably improved, humoral immunity, cellular immunity and mucosal immune responses of pregnant animals can be remarkably induced, maternal antibodies are generated, and the immune response efficiency of the pregnant animals is remarkably improved. The intestinal SIgA level of newborn animals is improved, and an effective technical means is provided for prevention and treatment of PED.

The invention discloses decapeptide with antioxidant activity, the amino acid sequence of the decapeptide is amino terminal-APKTLPWGPK-carboxyl terminal, and the decapeptide can be developed into an antioxidant.

The present invention relates to the fields of biomedicine and daily chemicals, and discloses a bioactive peptide having an amino acid sequence of ETRIL, or derivatives thereof having the same biological activity obtained by hydroxylating, carboxylating, carbonylating, methylating, acetylating, phosphorylating, esterifying, or glycosylating the amino acid side chain groups, the amino terminus, or the carboxyl terminus. The bioactive peptide has excellent anti-aging, antioxidant, and sunscreen skin care benefits, is homologous to yeast fermentation products, and can be used in pharmaceuticals, health products, foods, daily chemicals, and other fields, showing promising application prospects.

Polypeptides comprising at least one carboxy-terminal peptide (CTP) of chorionic gonadotrophin attached to the carboxy terminus but not to the amino terminus of a coagulation factor and polynucleotides encoding the same are disclosed. Pharmaceutical compositions comprising the polypeptides and polynucleotides of the invention and methods of using and producing same are also disclosed.

The application discloses a method for conformational signal peptide-mediated secretion and expression of heterologous proteins in Escherichia coli, and specifically comprises the following steps: simultaneously fusing at least two heterologous proteins to a conformational signal peptide (such as sfGFP) and constructing a heterologous recombinant protein expression vector, wherein the conformational signal peptide has a beta-barrel structure, and the fusion sites are at least two of the amino terminal end, the carboxyl terminal end and a plurality of loop rings of the conformational signal peptide; and then transforming the heterologous recombinant protein expression vector into Escherichia coli and culturing the obtained genetic engineering strain for expression. According to the method, the extracellular secretion and expression of the heterologous proteins in the Escherichia coli can be realized without the mediation of a classical signal peptide, the sfGFP as a fusion tag has no influence on the function of the target protein, and the corresponding target protein can be quickly expressed and purified; and the method expands the secretion and expression path of the protein, can realize the simultaneous expression of a plurality of heterologous proteins, and can effectively improve the expression amount of the target protein.

The invention relates to an anti-mouse IgG nano antibody and alkaline phosphatase fusion protein as well as a preparation method and application thereof. The anti-mouse IgG nano antibody and alkaline phosphatase fusion protein sequentially comprises an anti-mouse IgG nano antibody, a connecting peptide and shewanella alkaline phosphatase from an amino terminal to a carboxyl terminal. The fusion expression of the anti-mouse IgG nano antibody and shewanella alkaline phosphatase in escherichia coli is designed, so that various defects of a chemical labeling method can be avoided, a high-purity and high-specific activity enzyme-labeled antibody can be prepared, the preparation period of a second antibody can be shortened, the preparation process of the second antibody can be simplified, the stability of batches can be improved, and the dependence on experimental animals can be reduced. The prepared anti-mouse IgG nano antibody and alkaline phosphatase fusion protein can be applied to the field of immunodetection, such as a chemiluminescence immunoassay method, an enzyme-linked immunosorbent assay method, an enzymatic fluorescence immunoassay method, an immunoblotting method and the like.

The invention provides a nanopore probe and application thereof, the nanopore probe comprises a polypeptide, the carboxyl terminal of the polypeptide is connected with a steric hindrance tag, the amino terminal of the polypeptide is connected with polyaspartic acid, and the polypeptide has a restriction enzyme cutting site specifically recognized by target protease. The method can prolong the retention time of the polypeptide in the nanopore, enhance the signal discernibility and improve the signal stability, thereby realizing stable and high-precision quantitative detection of the protease activity.

The invention relates to a biotinylation modified amino acid probe, a preparation method and application thereof, and a method and application for identifying amino acid binding protein. The amino acid probe is amino acid with biotin modification at an amino terminal or a carboxyl terminal; or, the amino acid probe is amino acid with an azide group at the carboxyl end, the amino acid probe reacts with alkynyl-containing biotin under the catalysis of copper ions through a click chemical reaction in a cell to generate biotin-modified amino acid, an azide-modified amino acid probe library is supplemented, and the amino acid probe is combined with immunoprecipitation and mass spectrometry technologies, so that the biotin-modified amino acid is obtained. Proteins capable of being directly combined with amino acids can be identified, an important tool is provided for follow-up research of amino acid sensing proteins, and the method has important application value in identification of amino acid binding proteins, research of amino acid sensing mechanisms and functions and determination of drug targets or screening of lead compounds.

The invention discloses a multivalent expression sheep listeria vaccine for cervical cancer and a preparation method thereof, and the method comprises the following steps: on the basis of replacing sheep listeria LI [delta] ilo:: hly with hemolysin, constructing to obtain auxotrophic sheep listeria LI [delta] ilo:: hly [delta] dal [delta] dat; the fusion antigen is constructed by alternately arranging amino terminal and carboxyl terminal structural domains of four types of HPV6 / 11 / 16 / 18 E6 and E7 proteins and retaining an overlapping region of 16 amino acids at a connecting site; inserting the fusion antigen into a non-resistance plasmid pCW631 which carries a dal gene and replaces an Amp gene with an asd gene, so as to obtain a non-resistance plasmid pCW636; the resistance-free plasmid pCW636 is electrically transferred into the auxotroph sheep listeria monocytogenes LIdeltailo:: hly deltadal deltadat, and the sheep listeria monocytogenes cervical cancer vaccine is obtained. The cervical cancer vaccine provided by the invention can express the E6E7 protein in a multivalent manner, so that multiple protection on cervical cancer is realized.

The invention belongs to the field of chemical synthesis, and particularly relates to a chemical synthesis method of elastin-like polypeptide. The invention provides a chemical synthesis method of elastin-like, which comprises the following steps of: constructing a pentapeptide repetitive sequence (valine-proline-glycine-X-glycine, X is amino acid except proline, and the amino acid except glycine is respectively and independently selected from levoamino acid or dextroamino acid) of elastin-like polypeptide through liquid phase reaction; the elastin-like polypeptide can be prepared with the target molecular weight of 1000-500000 by adding amino groups and carboxyl groups into an organic solvent, forming orthogonal protection on the amino groups and the carboxyl groups, then selectively removing amino or carboxyl end protecting groups respectively, accurately coupling the amino groups and the carboxyl groups in a solution and repeating the deprotection and coupling processes for multiple times. The preparation method can realize accurate regulation and control of the molecular weight, sequence, chirality and topological structure of the elastin-like protein, improves the synthesis efficiency of the high-molecular-weight elastin-like polypeptide, has the advantages of large synthesis scale, low cost and the like, and has a wide application prospect.

The present inventors succeeded in improving the antibody constant region to have increased stability under acid conditions, reduced heterogeneity originated from disulfide bonds in the hinge region, reduced heterogeneity originated from the H chain C terminus, and increased stability at high concentrations as well as in discovering novel constant region sequences having reduced Fcγ receptor-binding, while minimizing the generation of novel T-cell epitope peptides. As a result, the present inventors successfully discovered antibody constant regions with improved physicochemical properties (stability and homogeneity), immunogenicity, safety, and pharmacokinetics.

Provided are a polypeptide, a polypeptide-modified magnetic bead, a preparation method therefor, and a use thereof. The C terminus of the polypeptide is modified with cysteine, and the amino acid sequence of the polypeptide is selected from one or more of SEQ ID NOs: 2-4. The polypeptide-modified magnetic bead comprises a magnetic bead, a linker unit, and a polypeptide, wherein the surface of the magnetic bead is modified with hydroxyl; the C terminus of the polypeptide is modified with cysteine, and the amino acid sequence of the polypeptide is selected from one or more of SEQ ID NOs: 1-4; and the end of one side of the linker unit is connected to the surface of the magnetic bead by means of a hydrogen bond, and the end of the other side of the linker unit is connected to the C terminus of the polypeptide by means of a thioether bond. The polypeptide can bind to a specific plasma protein in a targeted manner, especially a low-abundance plasma protein. When the obtained polypeptide-modified magnetic bead is used for plasma protein detection, the binding and detection of the low-abundance protein can be realized without separating high- and low-abundance proteins in plasma. The polypeptide-modified magnetic bead has universality.

The invention discloses a Pichia pastoris indirect surface display system based on a TRAP protein scaffold, which is characterized in that Sed1p protein from saccharomyces cerevisiae is used as anchoring protein, and tetrapeptide repeat affinity protein TRAP is fixed on the anchoring protein on the surface of a Pichia pastoris cell to obtain the Pichia pastoris indirect surface display system based on the TRAP protein scaffold. The target protein with the carboxyl end fused with the peptide MEEVV or the peptide MRRVW is indirectly displayed on the surface of the pichia pastoris cell through specific binding with the tetrapeptide repeat affinity protein TRAP. The xylanase and the beta-xylosidase are indirectly displayed on the surface of the pichia pastoris cell by utilizing the surface display system, so that the pichia pastoris surface display system has the catalytic capabilities of the xylanase and the beta-xylosidase, can perform a synergistic effect to degrade the zelkova xylan, and has a good application prospect.

The present invention discloses a recombinant humanized type XVII collagen, its expression vector and genetically engineered bacteria. In the present invention, 4 functional fragments with high activity are obtained through screening. After optimizing one of the fragments and combining them in a certain order, recombinant humanized type XVII collagen JYC1703 is obtained. Or after optimizing one of the fragments, combining them in a certain order and respectively fusing GPP sequences at the amino terminus and carboxyl terminus, recombinant humanized type XVII collagens JYC1701 and JYC1702 are obtained. After codon optimization, they are amplified and cloned into an expression vector. After enzymatic digestion and linearization, they are transformed into Pichia pastoris competent cells. High-copy recombinants are screened through G418 resistance gradient to obtain Pichia pastoris genetically engineered bacteria expressing recombinant humanized type XVII collagen. The recombinant humanized type XVII collagens JYC1701, JYC1702 and JYC1703 of the present invention exhibit excellent functions of promoting cell adhesion, cell proliferation and migration, and are suitable for preparing products such as skin care or medical devices.

A multi-specific antibody contains at least one binding site for a cell specific antigen and at least one binding site for a tumor-reactive lymphocyte antigen and a multi-specific antibody including an IgG antibody or fragment thereof that binds to a first antigen; and at least one scFv antibody that binds to a second antigen that is different from the first antigen and is linked to a C terminus of at least one light chain or heavy chain of said IgG antibody or fragment. The multi-specific antibody reversibly binds to at least one of the cell specific antigen and tumor-reactive lymphocyte antigen or the first antigen and the second antigen with a greater affinity at an aberrant condition than at a normal physiological condition. Conjugates of the multi-specific antibodies and methods for generating the multi-specific antibody are also provided.

This invention discloses a lipopeptide HIV membrane fusion inhibitor and its pharmaceutical use. The lipopeptide of this invention is of formula I: X1-EMTWEEWEK KVEELEKKIEELLK-X2-X3-X4-X5; or formula II: X1-ELTWEEWEKKVEELEKKIEELLK-X6-X7-X4-X5; wherein, X1 is an amino-terminal protecting group; X2 is a polypeptide sequence (EAAAK)n, A[(EAAAK)n]A, or (EP)n, where n represents the number of repetitions; X3 is Lys, Cys, Dap, Orn, Dab, Dah, or absent; X4 is a lipophilic compound group used to modify the C-terminus; X5 is a carboxyl-terminal protecting group; X6 is a polypeptide sequence KAEEQQKKNE; and X7 is Lys, Cys, Dap, Orn, Dab, or Dah. The lipopeptide of this invention has higher antiviral activity.

The invention provides a whey protein isolate source antioxidant polypeptide, the amino acid sequence of the whey protein isolate source antioxidant polypeptide is amino terminal-LVYPFPGPIP-carboxyl terminal, and the whey protein isolate source antioxidant polypeptide can be developed into an antioxidant.

The present disclosure relates to bispecific binding proteins comprising, from the amino terminal to the carboxy terminal, (a) a first moiety, which is an anti-BAFF antibody or antigen binding fragment thereof, and (b) a second moiety, which is an anti-MASP2scFv or truncated TACI polypeptide. Furthermore, the invention relates to nucleic acid molecules encoding such bispecific binding proteins, as well as vectors and host cells comprising such nucleic acid molecules. The invention further relates to a method for producing said bispecific binding proteins and to the use of said bispecific binding proteins in the treatment of autoimmune diseases.

The present invention discloses an artificial virus receptor and its application, which belongs to the field of biotechnology. The artificial virus receptor of the present invention includes modules A, B, C, D, E, F, G, and H from the amino terminus to the carboxyl terminus. Module A is a signal peptide of type I membrane protein; module B is a customized virus binding domain for capturing target viruses; modules C and E are linkers; module D is a spacer region for forming the skeleton of a customized artificial virus receptor to maintain a certain length outside the cell; module F is a transmembrane domain for realizing cell surface display of the artificial virus receptor; module G is an intracellular domain, which has the function of maintaining the stability of the artificial virus receptor; module H is a tag protein. The present invention paves the way for the rapid construction of in vitro and in vivo infection systems of viruses that are difficult to culture, especially high-risk coronaviruses, and for downstream prospective research, which will help to further advance the life cycle research of many difficult-to-culture viruses and promote the development of antiviral treatments and vaccines.

Disclosed herein, are peptides comprising from N terminus to C terminus, a) a peptide of interest, wherein the peptide of interest having 6 to 20 amino acid residues and a C -terminal residue that facilitates cyclization; b) a G macrocyclase recognition site; c) a linker; and d) G macrocyclase; wherein the peptide is linear, wherein the peptide of interest is covalently bonded to the G macrocyclase recognition site, wherein the G macrocyclase recognition site is bond to the linker, and wherein the linker is bonded to the G macrocyclase, and methods of making cyclized peptides and generating libraries of cyclized peptides.