23results about How to "Efficient secretion" patented technology

The present invention belongs to the field of biotechnology, and relates to a hybridoma cell strain secreting a monoclonal antibody of Clostridium perfringens CPA with neutralizing activity and its application. The present invention uses natural CPA as an immune antigen and a non-toxic CPA mutant as a coating antigen for the first time, and screens a hybridoma cell strain secreting a monoclonal antibody with neutralizing activity. The hybridoma cell strain can stably and efficiently secrete a monoclonal antibody with neutralizing activity, and can achieve large-scale batch production, thereby providing a material basis for the detection and treatment of CPA antigen / antibody in the later stage.

The invention relates to the field of biological medicines, in particular to an anti-CEACAM5 and CEACAM6 dual-targeting antibody and application thereof, the anti-CEACAM5 and CEACAM6 dual-targeting antibody comprises a heavy chain variable region, and the heavy chain variable region of the anti-CD66e antibody comprises the following technical characteristics: 1t; 1gt; comprising CDR-H1 with an amino acid sequence as shown in SEQ ID No. 1; lt; 2gt; comprising CDR-H2 with an amino acid sequence as shown in SEQ ID No.2; lt; 3gt; comprising CDR-H3 with an amino acid sequence as shown in SEQ ID No.3. The anti-CEACAM5 and CEACAM6 dual-targeting antibody provided by the invention has better affinity, the anti-CEACAM5 and CEACAM6 dual-targeting antibody is used as an antigen binding structural domain to construct a chimeric antigen receptor, the chimeric antigen receptor is used for preparing a CAR-T cell, and the CAR-T cell has killing activity on positive tumor cells of CEACAM5 and / or CEACAM6, can be effectively applied to immunotherapy, and has a good application prospect. The important significance is realized on the development of tumor treatment medicines.

The invention relates to the technical field of bacterial cellulose preparation, in particular to a composite flora for producing bacterial cellulose and a preparation method thereof, and the preparation method comprises the following steps: screening gluconobacter oxydans, obtaining culture medium preparation materials and preparation tools, preparing a slant culture medium set, preparing an enriched liquid culture medium set, and preparing composite flora mixed inoculation liquid. The method comprises the following steps: preparing a waste tobacco leaf leaching solution, extracting a proportional leaching solution from the waste tobacco leaf leaching solution according to a preset proportion, obtaining a mixed fermentation culture medium based on a preparation tool and the proportional leaching solution, and carrying out inoculation operation on a composite flora mixed inoculation solution by using the mixed fermentation culture medium to obtain a dry cellulose membrane; calculating the yield of the bacterial cellulose based on the dry cellulose membrane, and completing the preparation of the composite flora for producing the bacterial cellulose based on the yield of the bacterial cellulose. The method can be used for scientifically, efficiently and stably preparing the composite flora.

The invention provides bacillus licheniformis capable of producing amylase, protease and cellulase and application of the bacillus licheniformis. The bacillus licheniformis is bacillus licheniformis FJAT-27214 and is preserved in the China General Microbiological Culture Collection Center on August 6, 2025, the preservation address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No. 35536. The bacillus licheniformis disclosed by the invention can be used for simultaneously producing amylase, protease and cellulase at high yield; the bacillus subtilis has antibacterial activity to escherichia coli and has certain tolerance to artificial gastric juice, artificial intestinal juice and cholate, the survival rate in the artificial gastric juice is 58.11%, the survival rate in the artificial intestinal juice is 82.58%, and the survival rate in the cholate is 28.03%. When the strain fermentation liquor is used for pomace micro-storage, the content of crude protein and total sugar of pomace can be remarkably increased, meanwhile, the content of neutral detergent fiber and ammoniacal nitrogen is remarkably reduced, and an effective strain resource is provided for pomace micro-storage fermented feed.

The invention discloses a polypeptide module for promoting protein expression. The polypeptide module is polypeptide with an amino acid sequence as shown in SEQ ID NO: 2, wherein the polypeptide is formed by connecting a C end of a saccharomyces cerevisiae source signal peptide alpha-factor Pro section with two acidic residues EA; the composite signal peptide is formed by connecting a signal peptide OST1 from saccharomyces cerevisiae to the polypeptide module at the C end, the fusion expression of the composite signal peptide and the exogenous recombinant protein in saccharomycetes can improve the expression and secretion of the exogenous recombinant protein, and the composite signal peptide has popularization and application prospects.

The present application relates to the technical field of biological medicine, in particular to an anti-CEACAM5 antibody and application thereof, the anti-CEACAM5 antibody has the following technical features: the heavy chain variable region includes CDR-H1 with the amino acid sequence shown in SEQ ID No.1, CDR-H2 with the amino acid sequence shown in SEQ ID No.2 and CDR-H3 with the amino acid sequence shown in SEQ ID No.3.The anti-CEACAM5 antibody has high affinity, is used as an antigen binding domain to construct a chimeric antigen receptor, and utilizes the chimeric antigen receptor to prepare a CAR-T cell, the CAR-T cell has killing activity on tumor cells positive for CEACAM5, and after co-culturing with CEACAM5-positive cells, the CAR-T cell can efficiently secrete the cytokine IFN-gamma, the nanobody can be effectively applied to immunotherapy, and has important significance for developing tumor treatment drugs.

The present invention relates to a synthetic biology self-assembly vaccine production system and a method for producing a vaccine, wherein the system comprises a base sequence component capable of expressing an antigen, and the system is a sequence capable of enriching in host tissues and organs, expressing the antigen on the surface of a vesicle, and self-assembling into a vesicle complex, wherein the vesicle complex can induce an immune response. In the present invention, the base sequence component has the advantages of low production cost, stable physical and chemical properties, efficient antigen assembly and secretion process, fast response to pathogen mutation, strong immune response induced, high safety and protection, etc., providing a new type of vaccine.

The invention relates to the field of biological medicines, in particular to a CEA5 and NCA-90 bispecific antibody and application thereof, the CEA5 and NCA-90 bispecific antibody comprises a heavy chain variable region, and the heavy chain variable region of the anti-CD66e antibody comprises the following technical characteristics: 1t; 1gt; comprising CDR-H1 of which the amino acid sequence is shown as SEQ ID No.1 or 2; lt; 2gt; comprising CDR-H2 with an amino acid sequence as shown in SEQ ID No.3; lt; 3gt; comprising CDR-H3 with an amino acid sequence as shown in SEQ ID No.4. The CEA5 and NCA-90 bispecific antibody provided by the invention has relatively good affinity, the CEA5 and NCA-90 bispecific antibody is used as an antigen binding structural domain to construct a chimeric antigen receptor, the chimeric antigen receptor is used for preparing a CAR-T cell, and the CAR-T cell has killing activity on CEA5 and / or NCA-90 positive tumor cells, can be effectively applied to immunotherapy, and has a good application prospect. The important significance is realized on the development of tumor treatment medicines.

The application relates to the technical field of bioengineering, and particularly discloses a recombinant human collagen XV and a preparation method and application thereof. The amino acid sequence of the recombinant human collagen XV is shown as SEQ ID NO. 1. The recombinant human collagen XV is derived from human collagen, and has no rejection and sensitization risks. Experiments show that the protein has good effects of promoting cell proliferation and promoting cell migration, and has good biocompatibility, and can be applied as a safe and effective biomedical material.

The application belongs to the technical field of biology, and discloses a hybridoma cell strain 2F8 secreting a calmodulin 2 monoclonal antibody, a specific monoclonal antibody, and preparation and application. The hybridoma cell strain 2F8 secreting the calmodulin 2 monoclonal antibody is classified and named as 2F8, and is preserved in the China General Microbiological Culture Collection Center on April 9, 2025, with a preservation number of CGMCC No.46343. The application also provides the calmodulin 2 monoclonal antibody secreted by the hybridoma cell strain 2F8 and a polynucleotide encoding the calmodulin 2 monoclonal antibody. The antibody has specific recognition ability for the calmodulin 2, and is suitable for immunological detection, in particular, Western-blot and immunohistochemical detection. The antibody can be used for preparing a reagent, a kit or other immunological detection product for detecting the calmodulin 2, and has practical application value.

The present invention relates to a thermophilic tennis beta-mannanase mutant with improved thermal stability and its application, a mutant with improved thermal stability is obtained by random mutagenesis and expressed in Bacillus subtilis. The results show that the optimum temperature of the β-mannanase mutant is 80 ° C, the optimum pH is 5.0, the shake flask fermentation is 4 days, and the konjac gum is used as the substrate, which is 4.1% higher than the wild-type enzyme activity; its half-life at 80 ° C is 5 h, which is 2 h more than the wild-type, and the enzyme is completely inactivated for more than 48 h. In addition, the ability of the β-mannanase mutant to hydrolyze locust bean gum and guar gum is 10.2% and 2.0% higher than the wild-type, respectively. The enzyme has shown good application prospects in the fields of oil extraction and animal breeding.

The invention discloses an anti-human CD47 monoclonal antibody which comprises a heavy chain variable region and a light chain variable region, and a complementarity determining region of the heavy chain variable region comprises CDRH1 with an amino acid sequence as shown in SEQ ID No.1; the complementarity determining region of the heavy chain variable region comprises CDRH2 of which the amino acid sequence is shown as SEQ ID No.2; the complementarity determining region of the heavy chain variable region comprises CDRH3 of which the amino acid sequence is shown as SEQ ID No.3; the complementarity determining region of the light chain variable region comprises CDRL1 of which the amino acid sequence is as shown in SEQ ID No.4; the complementarity determining region of the light chain variable region comprises an amino acid sequence CDRL2 which is KVS; the complementarity determining region of the light chain variable region comprises CDRL3 of which the amino acid sequence is as shown in SEQ ID No.5. The invention also discloses a preparation method of the anti-human CD47 monoclonal antibody and an application of the anti-human CD47 monoclonal antibody in CD47 protein immunodetection.

This invention relates to the fields of microbial fermentation and genetic engineering technology, specifically disclosing a method for preparing human type III collagen fragments. Based on Pichia pastoris, this invention constructs a multidimensional synergistic expression system of "molecular chaperone co-expression - precursor supply - osmotic pressure adaptation regulation," reducing the structural instability of collagen fragments during secretion and post-processing, and constructing an expression system suitable for stable and efficient secretion of collagen fragments. The method described in this invention can achieve efficient extracellular secretory expression of human type III collagen fragments, and the obtained protein has high structural uniformity and stability, with a protein yield of up to 6.55 g / L under 5 L fermenter conditions. The method described in this invention is suitable for the large-scale preparation of human type III collagen fragments and provides a new technical solution for obtaining collagen products with high structural uniformity and good functional activity.

The invention discloses a humic substance reinforced hydroxyapatite-bacillus subtilis semi-artificial photosynthetic material and a preparation method and application thereof.The preparation method comprises the steps that firstly, hydroxyapatite microspheres are subjected to Fe2O3 doping, the Fe2O3 doped hydroxyapatite microspheres are combined with bacillus subtilis, and the hydroxyapatite-bacillus subtilis semi-artificial photosynthetic material is obtained; and the semi-artificial photosynthetic material of the bacillus subtilis-Fe2O3 doped hydroxyapatite microspheres is obtained. And then, adding the humic substance into the hydroxyapatite-bacillus subtilis semi-artificial photosynthetic system, so as to obtain the humic substance reinforced hydroxyapatite-bacillus subtilis semi-artificial photosynthetic material. The raw materials of the prepared material are easy to obtain, the method is simple, and when the material is applied to enhancing the activity of the antioxidant enzyme of the bacillus subtilis, the cost of producing the antioxidant enzyme by the bacillus subtilis is expected to be reduced; meanwhile, the bacillus subtilis can be used for fixing CO2 through photocatalytic reduction, carbon emission and the like in the aerobic fermentation process of the bacillus subtilis are expected to be reduced, and economic benefits and environmental benefits are achieved.

The present invention discloses a humanized anti-human γδTCR monoclonal antibody and its encoding gene, preparation method and application, which belongs to the humanized monoclonal antibody preparation technology field in the biotechnology field. The heavy chain variable region of the humanized anti-human γδTCR monoclonal antibody provided by the present invention comprises the amino acid sequence shown in SEQ ID NO:4 or SEQ ID NO:5, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:7. The humanized anti-human γδTCR monoclonal antibody can stimulate and amplify peripheral blood γδT cells in vitro, and the obtained γδT cells can secrete a variety of cytokines and can effectively kill a variety of tumor cells.

The application provides a kind of recombinant hydroxylated small molecule collagen and its expression system and preparation method, belong to the field of bioengineering technology;The application utilizes synthetic biology, genetic engineering and biotechnology to co-express small molecule collagen and P4H, then carries out amino acid modification, hydroxylation is carried out at G-X-Y theory hydroxylation site, post-translational modification is carried out, and the recombinant hydroxylated small molecule collagen is obtained;The recombinant hydroxylated small molecule collagen of the application is stable in molecular structure by amino acid post-expression modification, and is more controllable in comparison with small molecule self-assembly after expression, which can balance transdermal absorption and biological structure stability;The recombinant hydroxylated small molecule collagen can realize industrial production, and has good application in the fields of drugs, medical devices, biomaterials, tissue engineering products, cosmetics or health products and the like.

The invention belongs to the technical field of biology, and particularly relates to a hybridoma cell strain capable of secreting a clostridium perfringens ETX monoclonal antibody with neutralizing activity and application of the hybridoma cell strain. On the basis of natural toxin, an indirect ELISA (enzyme-linked immunosorbent assay) detection method is established by utilizing a non-toxic ETX single amino acid site protein mutant, and a series of hybridoma cell strains secreting an anti-clostridium perfringens ETX monoclonal antibody are screened; the hybridoma cell strain can stably and efficiently secrete a monoclonal antibody with neutralizing activity and a monoclonal antibody without neutralizing resistance respectively, and large-scale batch production can be realized. According to the present invention, the clostridium perfringens ETX monoclonal antibody is firstly utilized to establish the ETX antigen ELISA detection method, and the method has characteristics of simple sample operation, low cost, rapid reaction, strong specificity and the like, can provide the reference for the diagnosis of clostridium perfringens, and provides the basis for the research of the related vaccine potency test substitution method.

The application discloses a recombinant plasmid for expressing salt-tolerant protease, a yeast engineering bacterium, a fermentation medium thereof and application. Key elements for heterologous expression of the salt-tolerant protease are confirmed, and the recombinant plasmid for expressing the salt-tolerant protease and the yeast engineering bacterium are constructed. The long-term salt-tolerance level and SDS tolerance of the salt-tolerant protease expressed by the yeast engineering bacterium are significantly improved, and the salt-tolerant protease can effectively and significantly degrade soybean protein isolate under high-temperature and high-salt conditions, so that the salt-tolerant protease has important application value for efficient catalysis of protein substrates in traditional high-salt fermentation industry. The application also realizes efficient secretion of the salt-tolerant protease by optimizing the concentration of enzyme cofactors. Furthermore, electrophoretically pure grade recombinant salt-tolerant protease is obtained through one-step separation and purification, and the purification process is simple and efficient.

The invention belongs to the technical field of biology, and particularly relates to a hybridoma cell strain capable of secreting a clostridium perfringens ETX monoclonal antibody with neutralizing activity and application of the hybridoma cell strain. On the basis of selecting natural toxins, an indirect ELISA detection method is established by utilizing nontoxic ETX single amino acid site protein mutants, a series of hybridoma cell strains secreting anti-clostridium perfringens ETX monoclonal antibodies are screened, and the hybridoma cell strains can stably and efficiently secrete the monoclonal antibodies with neutralizing activity. According to the present invention, the clostridium perfringens ETX monoclonal antibody with neutralization resistance is combined with the ETX tetraamino acid mutant to establish the ETX antibody blocking ELISA detection method, and the method can reduce the potential safety hazard of the ETX natural toxicity and the potential safety hazard of the single amino acid mutant amino acid reverse mutation; the kit also has the characteristics of simplicity and convenience in operation, low cost, quick reaction, strong specificity and the like, and can provide reference for diagnosis of clostridium perfringens disease.

The present invention discloses a recombinant Bacillus subtilis producing walnut active peptides, a construction method and an application, and belongs to the field of bioengineering technology. The recombinant Bacillus subtilis producing walnut active peptides comprises a recombinant plasmid and an expression host; the recombinant plasmid is formed by introducing the HHHHHHK polypeptide coding sequence at the N-terminus of the active peptide TWLPLPR or the active peptide mutant TWLPYPR tandem sequence, and then homologously recombine into the expression plasmid; the expression host is Bacillus subtilis. The present invention adopts the strategy of tandem expression and signal peptide and host optimization to achieve efficient secretory expression of walnut active peptides and mutants thereof in food-grade hosts. The present invention lays a certain foundation for the efficient preparation of small molecule active short peptides and is suitable for industrial production and application.

The invention relates to the technical field of veterinary products, in particular to an artificially synthesized bovine rotavirus VP7-M protein mutant, a vaccine and application. According to the designed bovine rotavirus VP7-M mutant, disulfide bonds and hydrophobic hole filling mutation are introduced, five amino acid mutations are designed in an original protein amino acid sequence, and then a CHO cell line capable of stably expressing the bovine rotavirus VP7 protein mutant is constructed; the cell line can efficiently secrete and express VP7-M mutant protein, the conformation of the VP7-M mutant protein is similar to that of natural protein, the secreted and expressed VP7-M mutant protein is subjected to one-step affinity chromatography purification and prepared into the subunit vaccine, the immunogenicity of the VP7-M mutant protein is evaluated by immunizing a mouse, and animal experiment results show that the VP7-M subunit vaccine is good in safety and has good application prospects. The bovine rotavirus subunit vaccine has good immunogenicity and has a protection effect similar to or even better than that of a traditional inactivated vaccine, so that a foundation is laid for further preparation of the bovine rotavirus subunit vaccine.

The invention provides bacillus subtillis capable of producing amylase, protease and cellulase and application of the bacillus subtillis, the bacillus subtillis is bacillus subtillis FJAT-10612, the bacillus subtillis is preserved in China General Microbiological Culture Collection Center on August 6, 2025, the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No.35537. The amylase production activity of the bacillus subtillis disclosed by the invention is 166.12 U / mL, the cellulase production activity of the bacillus subtillis is 88.05 U / mL, and the protease production activity of the bacillus subtillis is 18.36 U / mL; certain tolerance to artificial gastric juice, artificial intestinal juice and cholate is achieved; when the strain fermentation liquor is used for pomace micro-storage, the content of pomace crude protein and total sugar can be remarkably increased, the cellulase activity is improved, meanwhile, the content of crude fibers, neutral detergent fibers, lignin and ammoniacal nitrogen is remarkably reduced, and an effective strain resource is provided for pomace micro-storage fermented feed.

The application belongs to the technical field of biocatalytic synthesis of steroid drugs, and particularly relates to a dehydrogenase, a gene, an engineering bacterium and application, the amino acid sequence of the dehydrogenase is SEQ ID NO. 1 or SEQ ID NO. 2, the dehydrogenase has high dehydrogenation activity, high stereoselectivity and strong regioselectivity, and no other conformation impurities are generated; the engineering bacterium can efficiently secrete the dehydrogenase, and effectively improves the conversion efficiency of a substrate, especially for a substrate 6-methy format (11beta, 17alpha-dihydroxy-6alpha-methylpregna-1, 4-diene-3, 20-diketone), which is low in cost and has significant industrialization potential; the dehydrogenase mutant has clear physicochemical properties, and the method of whole cell catalysis is used, so that the cumbersome step of enzyme purification is omitted, and the cost is reduced and the operation is simple.