256 results about "Cell Membrane Proteins" patented technology

The invention discloses a fermentation method of recombinant bacillus subtilis for expressing vibrio parahaemolyticus outer membrane protein, which comprises the following steps: inoculating a recombinant bacillus subtilis strain into a liquid culture medium for activation and amplification to obtain a seed solution; transferring the seed solution into a fermentation tank according to the inoculum size of 5%-10%, controlling the temperature to be 35-37 DEG C and the pH to be 6.8-7.2, introducing sterile air and stirring, and monitoring the thallus concentration OD600 value in real time; when the OD600 value reaches 35-38, 10% of an inducer is added for pre-induction; when the OD600 value reaches 40-45, the remaining 90% of the inducer is added for main induction, and the real-time dissolved oxygen value is collected; when the real-time dissolved oxygen value is continuously higher than a first preset threshold value, the flow acceleration rate of the inducer is increased, the fermentation temperature is increased at the same time or step by step, and the increase amplitude of the dissolved oxygen value is in positive correlation with the flow acceleration rate and the temperature increase amplitude; when the OD600 value is stabilized in a preset interval, it is judged that fermentation is completed, and the recombinant bacillus subtilis thalli are obtained. The yield and quality of the vibrio parahaemolyticus outer membrane protein can be effectively improved.

The invention discloses a cell membrane protein replacement type oncolytic virus vector and application thereof. The cell membrane protein replacement type oncolytic virus vector is rhabdoviridae virus, and a nucleotide sequence for coding G protein in a genome of the rhabdoviridae virus is replaced by a nucleotide sequence for coding an antibody and a nucleotide sequence for coding a spike protein truncation of coronavirus. The invention also discloses a construction method of the cell membrane protein replacement type oncolytic virus vector for expressing the antibody, the non-replicated virus vector is used for expressing the antibody sequence for the first time, and meanwhile, the novel coronavirus cell membrane protein is embedded into the virus surface, so that the cell membrane protein replacement type oncolytic virus vector can be rapidly produced in a suspension cell in a large scale; through removal of virus cell membrane protein genes, the virus cell membrane protein genes cannot be continuously replicated in vivo, so that the safety of the virus cell membrane protein genes is ensured, the tumor immunosuppression condition is improved, and an organism can be stimulated to generate a neutralizing antibody for resisting new coronavirus while tumor cells are killed.

The invention belongs to the technical field of biological medicine, and particularly relates to an engineered cell microvesicle and a delivery system based on the engineered cell microvesicle, the system realizes efficient preparation of 1-5 [mu] m cell microvesicles, and the cell microvesicles have a large space volume and can be used for preparing the cell microvesicles. The carrier can be used for loading and delivery of target protein, polypeptide and recombinase which are specifically expressed in mother cells. The system transfects mother cells through lentivirus transfection or plasmid transfection to further produce cell microvesicles, and the microvesicles can load more goods and inherit membrane proteins of the mother cells, and can also effectively load intracellular proteins to realize effective delivery. By virtue of good structural stability, high immunogenicity and excellent biocompatibility, the cell microvesicle reduces systematic toxic and side effects of a traditional carrier, is expected to become an effective drug delivery system, and has great application potential in the field of gene therapy.

The invention discloses a breast milk fat globule membrane protein simulation degree evaluation method based on multi-dimensional similarity weighted fusion, and the method comprises the steps: constructing a breast milk MFGM proteomics database, and obtaining the proteomics data of a to-be-evaluated sample; the method comprises the following steps: obtaining a reference matrix through row screening and column screening based on a breast milk MFGM proteomics database, performing logarithmic transformation on the reference matrix, calculating a mean value and a standard deviation of each feature in the reference matrix, and constructing a Z-score matrix and a missing mask matrix corresponding to the reference matrix; calculating abundance distribution similarity and missing mode similarity of the to-be-evaluated sample; setting a shrinkage coefficient based on the actual overlapping feature number, screening a core protein set according to a set threshold value, and comparing to obtain the missing MFGM protein type of the to-be-detected sample; and constructing a Top-K neighbor center, calculating the feature deviation contribution and abundance difference of the to-be-evaluated sample, and outputting key defect proteins and formula optimization suggestions according to preset feature importance indexes. And the similarity simulation accuracy is improved.

The invention provides an organoid membrane protein and intracellular protein immunofluorescence in-situ co-staining method, which comprises the following steps of: sealing and fixing organoid glue drops, adding a mixed solution of membrane protein primary antibody and a Dune phosphate buffer solution containing bovine serum albumin for incubation, performing membrane permeation by using a permeation agent, sealing and washing, and drying to obtain the organoid membrane protein and intracellular protein immunofluorescence in-situ co-staining product. Adding a mixed solution of an intracellular protein primary antibody and a Dune phosphate buffer solution containing bovine serum albumin into the cell plate where the transparent organ-like glue drops are located, incubating and washing, then adding a mixed solution of a membrane protein and a fluorescent secondary antibody of the corresponding species of the intracellular protein, incubating and washing in a dark place, and finally obtaining the membrane protein / intracellular protein fluorescent antibody. And finally, adding a mixed solution of DAPI and a Duncheng phosphate buffer solution, and incubating and washing in a dark place again to obtain the membrane protein and intracellular protein immunofluorescence in-situ co-dyed organoid. According to the method, high-quality and high-fidelity membrane protein and intracellular protein in-situ co-dyeing is successfully realized, the integrity of a three-dimensional structure and the high efficiency of an experimental process are taken into consideration, and the method is convenient to operate, high in repeatability and good in universality.

The application discloses a Vibrio harveyi oral vaccine and a preparation method and application thereof. The application uses a lactococcus lactis NZ9000 expression system, modifies transformation parameters, effectively expresses Vibrio harveyi OmpU protein, constructs a lactococcus lactis (NZ9000) engineering bacterium expressing Vibrio harveyi outer membrane protein (OmpU), develops an oral vaccine against Vibrio harveyi infection, aims to reduce the infection of economic animals in seawater aquaculture by oral administration, provides a new scheme for dealing with the harm of Vibrio harveyi infection and antibiotic resistance and the like, promotes the breeding of fry and fingerlings, and promotes the development of biological agriculture and related industries. The strategy is expected to overcome the problems of traditional injection vaccines, such as inconvenient operation and great stress, and can provide a new technical scheme for reducing the dependence of the aquaculture industry on antibiotics and promoting the establishment of a green and healthy breeding mode.

This invention discloses an antigen and vaccine preparation based on dominant Brucella epitopes, and their application, belonging to the field of recombinant subunit vaccine technology. The antigen is composed of CTL epitopes, HTL epitopes, and B-cell epitopes tandemly, and its amino acid sequence is shown in SEQ ID NO.1. Based on screened dominant CTL epitopes, HTL epitopes, and B-cell epitopes of the Omp25 and Omp31 outer membrane proteins, this invention constructs a novel antigen fusion polypeptide. The vaccine prepared based on this polypeptide effectively activates a significant dual immune response, enhancing cellular immunity while inducing the body to produce high levels of specific antibodies. It also provides good protection against organ damage caused by bacterial infection, offering better immune response and protective efficacy against Brucella, laying the foundation for the development of Brucella recombinant subunit vaccines.

A method for predicting vertigo targets using membrane protein-specific weighted interaction enhancement belongs to the interdisciplinary field of bioinformatics and machine learning. This invention uses vertigo membrane protein targets as the core adaptation object, constructing a complete technical chain of "membrane protein-specific multi-dimensional feature input → MRMR feature screening → membrane protein-specific feature correlation weighting – cross-view interaction enhancement fusion → normalization processing → SVM classification prediction." All parameters are fixed and optimized throughout the process, requiring no adjustment based on test data. Through specific embodiment testing and multi-dimensional verification, it is proven that the method's prediction results are realistic, effective, and stable. The prediction accuracy and efficiency are significantly better than existing mainstream methods, and it is currently one of the few dedicated prediction methods specifically designed for vertigo membrane protein targets, filling a gap in existing technology.

This invention discloses an enhanced CAR-T cell (CARTAC) capable of secreting a targeted membrane protein degrader and its applications. While retaining specific killing function, this cell can expand locally in tumors and continuously secrete the bispecific adaptor molecule scTAC. scTAC can recruit E3 ubiquitin ligases, mediate ubiquitination of tumor cell surface membrane proteins (such as PD-L1), and achieve targeted degradation via the lysosomal pathway, effectively reversing the immunosuppressive microenvironment, overcoming antigen heterogeneity, and eliminating antigen-negative tumor cells through the bystander effect. This invention also utilizes the efficient endocytosis properties of EGFR to design scTAC-dual, which can further enhance degradation efficiency. In various lung cancer mouse models, CARTAC-dual exhibited excellent tumor infiltration capacity, low exhaustion levels, and sustained antitumor activity, without systemic toxicity. This invention integrates the targeted killing and protein degradation delivery functions of CAR-T cells, not only enhancing the control effect on solid tumors but also possessing target scalability, making it suitable for the universal upgrading of various CAR-T products.

The invention belongs to the technical field of molecular biology, and discloses a truncated outer membrane protein of klebsiella pneumoniae and application thereof. The invention provides an outer membrane protein Lol B of klebsiella pneumoniae, which can be used for immunizing to induce a protective antibody aiming at various serotypes of klebsiella pneumoniae and killing the klebsiella pneumoniae. Specifically, the outer membrane protein Lol B immune serum can efficiently kill Klebsiella pneumoniae, and when a mouse immunized by the outer membrane protein Lol B immune serum is attacked by K1, K2 and K6 serotype Klebsiella pneumoniae, the mouse can be completely protected to survive, and the trend of weight loss of the mouse is remarkably reduced. Therefore, the outer membrane protein Lol B provided by the invention has a clinical application value for preventing and treating klebsiella pneumoniae infection.

The invention discloses application of rutin hydrate in preventing and treating repeated embryo planting failure, and belongs to the technical field of assisted reproduction. It is found for the first time that CD36 is a key membrane protein for lipid transport and uptake of EECs, rutin hydrate competitively inhibits combination of CD36 and natural ligands of CD36, abnormal lipid excessive uptake is remarkably blocked, lipid accumulation and lipid toxicity damage in cells are effectively relieved, and the effect of inhibiting EECs lipid transport and uptake is achieved. And finally, the endometrial receptivity defect related to RIF is reversed, and the embryo adhesion and implantation capability is remarkably improved. Therefore, rutin hydrate provides a brand new strategy for developing targeted therapy drugs for RIF, especially for lipid metabolism disorder type endometrial receptivity badness, and has remarkable clinical application and transformation prospects.

To provide a high-affinity monoclonal antibody for a His-tag added to the C terminus of membrane protein.SOLUTION: The present invention provides a monoclonal antibody for a histidine tag (His-tag) or an antigen-binding fragment thereof, wherein the monoclonal antibody recognizes, as an epitope, a His-tag added to the C terminus of a membrane protein.SELECTED DRAWING: None

The invention discloses an engineered protein and application thereof in mediating non-ubiquitination degradation, and belongs to the technical field of biological medicine. The structural domains CATCH1 and CATCH2 of the MIDN protein and the region between the structural domains CATCH1 and CATCH2 are modified for the first time, the modification mode comprises the steps that the structural domains CATCH1 and CATCH2 and the region between the structural domains CATCH1 and CATCH2 are replaced with antibodies or polypeptides, then structure or sequence optimization is conducted on the basis, and the obtained MIDN engineered protein not only retains the proteasome binding capacity of MIDN, but also has the advantages of being capable of improving the protein quality and the like. According to the present invention, the MIDN-based target protein degradation system has the MIDN-based target protein degradation system, can expand the target range to the membrane protein or the cytoplasm protein, can improve the degradation efficiency of the MIDN-based target protein degradation system, further has the antibody-mediated target protein specificity so as to achieve the pathological / physiological protein distinguishing effect, and does not have the obvious toxicity; therefore, the engineered protein provided by the invention has a good application prospect.

The invention discloses a cell membrane in-situ drug membrane protein target screening method based on variable configuration DNA, and belongs to the technical field of biological medicine. Two pairs of functionalized nucleic acid structures are constructed, the first pair is S1S2 semi-complementary DNA double strands, the second pair is S3S4 DNA-RNA hybrid double strands, and the S1S2 structure is covalently coupled to a non-natural amino acid site-directed modified membrane protein through a click chemical reaction in a living cell to realize tagging of the membrane protein; when the candidate drug coupled in S3 interacts with the membrane protein, triggering an allosteric response system to release the tagged chain in S1; the released DNA tag is subjected to PCR amplification and sequencing, and accurate identification and analysis of a membrane protein target are realized according to a tag sequence. The invention provides a high-specificity and non-in-vitro drug membrane protein target screening technology. According to the technology, active compound discovery or accurate screening of drug membrane protein targets in a living cell in-situ environment can be realized.

Provided are a recombinant Escherichia coli with improved ability to secrete extracellular endoglucanase (BcsZ) and a construction method therefor. The recombinant Escherichia coli is constructed by using Escherichia coli JBZ-DH5α as a starting strain and inserting a membrane protein coding gene. The membrane protein gene is one of the following coding genes: a coding gene of zinc transporter permease znuB, a coding gene of DUF1449 family inner membrane protein ylaC, and a coding gene of UPF0410 family inner membrane protein ymgE.

Embodiments of the invention disclosed herein involve the selective engineering of Sindbis virus so as to generate cells that make enveloped virus-like particles that contain therapeutic mRNAs, and whose membrane proteins have been mutated to serve as modular binding sites for cell-targeting ligands.

The invention discloses application of sEVs membrane protein detection in breast cancer drug resistance early warning products, and belongs to the field of biomedicine. A plurality of breast cancer drug resistance associated sEVs membrane proteins are selected to form the combined marker, so that the detection accuracy can be effectively improved. By applying the antibody, the primer and the probe designed by the invention and combining the design of four-layer cascade amplification and multi-target conjoint analysis, four-round signal amplification can be completed, the limitation of a traditional single-target detection method is effectively broken through, the detection sensitivity is remarkably improved, the detection time is shortened, the detection cost is reduced, and the possibility is provided for clinical large-scale screening.

The application belongs to the technical field of biological medicine, and particularly relates to a composite material containing a local anesthetic and a preparation method thereof. The composite material containing the local anesthetic comprises the following raw materials in parts by weight: bupivacaine hydrochloride 10-15 parts, a light-heat conversion agent 4-8 parts, delta-cadinene 0.5-1 part, a penetration enhancer 1-3 parts, and glycerol 2-5 parts. The light-heat conversion agent and the bupivacaine hydrochloride are added in combination, so that the discomfort caused by photothermal therapy is relieved. The delta-cadinene and the bupivacaine hydrochloride are combined, so that the analgesic effect is improved, the analgesic time is prolonged, the pain caused by frequent dressing change is avoided, and the toxic side effects caused by the accumulation of blood drug concentration are avoided. In addition, the penetration enhancer composed of egg membrane protein peptide and polylysine is introduced, so that the absorption of the effective components by the skin is further promoted, and the analgesic effect is quickly exerted.

The invention belongs to the technical field of biology, and relates to application of a ZmCASPL11 gene in improving heat resistance of plants. The ZmCASPL11 gene encodes a protein of a membrane protein (CASPL) family related to a Kjeldahl band structure, the gene expression level of the ZmCASPL11 gene under high-temperature stress is remarkably increased, and a mutant zmcaspl11 of the gene shows a high-temperature sensitive phenotype. The ZmCASPL11 gene disclosed by the invention is transformed into arabidopsis thaliana and is subjected to functional verification, and compared with wild type arabidopsis thaliana, the tolerance of an arabidopsis thaliana plant over-expressed with the ZmCASPL11 gene to heat stress is remarkably enhanced. According to the invention, more theoretical bases are provided for corn heat resistance research, and gene resources and biotechnological means are provided for subsequent cultivation and germplasm resource innovation of new heat-resistant corn strains.

The application discloses a Mycoplasma bovum adhesion protein FtsZ and application thereof. The nucleic acid sequence of the Mycoplasma bovum adhesion protein FtsZ is shown as SEQ ID NO. 1. The application also discloses a recombinant plasmid pET-30a-ftsZ and an E. coli containing the recombinant plasmid pET-30a-ftsZ. ftsZ The recombinant protein rFtsZ has the advantages of being capable of specifically combining with EBL cell membrane protein, being capable of combining with extracellular matrix components (fibronectin, fibronectin, laminin and type IV collagen), having the direct adhesion host cell effect, having good antigenicity, being capable of producing high-level antibodies, being capable of providing good immune protection effect, and providing a new target and thought for elucidating the pathogenic mechanism of the Mycoplasma bovum and developing a new vaccine and medicine.

The invention relates to application of IGFBP7 in clinical diagnosis of colorectal cancer and application of IGFBP7 as a drug target. The research shows that the expression quantity of IGFBP7 secreted by tumor endothelial cells is continuously reduced in the colorectal tumor formation process related to inflammation. Functional analysis proves that the endothelial-derived IGFBP7 is secreted into a tumor microenvironment and plays a role in resisting angiogenesis. In addition, the CRC cells can actively ingest the vesicles containing the IGFBP7. The IGFBP7 reduces the activity of a TGF-beta1 signal channel by inhibiting the expression of EGR1, thereby inhibiting the proliferation and migration of tumor cells. What is worthy of attention is that VAPA (Vesicle-associated Membrane Protein Associated Protein A) is identified to be a key factor for mediating IGFBP7 vesicles to be transferred to lysosome, and VAPA promotes the degradation of IGFBP7, so that the cancer suppression function of the IGFBP7 is weakened. The endothelial cell-derived IGFBP7 inhibits the progress of CRC (cyclic redundancy check) through an EGR1 / TGF-beta 1 pathway, and VAPA-mediated lysosome degradation limits the cancer inhibition effect of the endothelial cell-derived IGFBP7. The discovery emphasizes that the endothelial cell-derived IGFBP7 is a promising therapeutic target in the field of colorectal cancer.

This invention discloses an IGF2 fusion antibody, its construction method, and its applications. Using a PD-L1 antibody as a template, the IGF2 domain is fused monovalently or bivalently to the heavy or light chain end of the PD-L1 antibody, self-assembling to form four conformations of fusion antibodies: Type A, Type A-GS, Type B, or Type B-Fab. By constructing IGF2 fusion antibodies with different conformations, the optimal IGF2 fusion antibody conformation is obtained through affinity activity and protein degradation screening. Specifically, the IGF2 of the fusion antibody exhibits nanomolar affinity activity with the IGF2 receptor on the cell surface, achieving efficient degradation of various cell surface membrane proteins (such as PD-L1, HER2, EGFR, and GPC3). This invention provides a new technological platform for membrane protein degradation.

Implementations of an anti-tick vaccine composition may include one or more conserved tick proteins from a tick species; and one or more conserved outer membrane proteins of an endosymbiont of the tick species.

Provided herein are cell-based assays for measuring the immunomodulation of therapeutic biologics using immobilized recombinant proteins. Specifically, the present disclosure relates to a method of assessing and / or determining a biomolecule-induced immune response, comprising: a) immobilizing a recombinant membrane protein or a fragment comprising an extracellular domain thereof on a support; b) contacting the immobilized protein or fragment with (i) a biomolecule, such as an antibody, and (ii) effector cells, and c) detecting activation of the effector cells to assess and / or determine an immune response induced by the biomolecule, such as ADCC, ADCP.

The invention provides a regulatory T cell with improved phenotype and functional stability and a preparation method and application thereof. The regulatory T cell disclosed by the invention is co-expressed by membrane protein GARP and TGF-beta, and specifically, the TGF-beta comprises a C33S point mutation precursor TGF-beta 1; further, the regulatory T cell co-expresses a cytokine and a sorting marker, specifically, the cytokine comprises IL-10, and the sorting marker comprises a truncated nerve growth factor receptor tNGFR. The regulatory T cell has more stable phenotype and function stability in in-vivo and in-vitro environments, has important enlightenment value for improving the function stability problem of the regulatory T cell in an inflammatory environment, and has wide application prospects in the fields of treatment of autoimmune diseases, intervention of organ transplantation rejection and the like.

The invention belongs to the technical field of nano biological materials, and particularly relates to a preparation method of cell membrane coated nanoparticles based on solvent equilibrium and sulfydryl anchoring. According to the method, sulfydryl-containing lipid is doped into a membrane system, so that the sulfydryl-containing lipid and the surface of a substrate form a covalent anchor bond to enhance the membrane-substrate binding force, and a small amount of organic solvent is added to improve the fluidity of the lipid and promote spontaneous fusion and uniform spreading of a membrane layer on the surfaces of particles. And centrifuging and washing to obtain the cell membrane nanoparticles with completely coated surfaces. According to the method, high-energy ultrasonic or extrusion treatment is not needed, stable coating can be realized on the premise of maintaining complete composition of membrane protein and lipid, and the adhesion stability of a membrane layer in a high-salt and shearing environment is remarkably improved. The method is suitable for various nano substrates and different morphological structures, and the obtained particles show higher antibody titer and excellent safety in an anti-alpha-hemolysin vaccine model. The invention provides a mild and efficient bionic nano material construction strategy with wide applicability.

The invention discloses a functionalized microarray pore plate, a preparation method thereof and application of the functionalized microarray pore plate in membrane protein ligand screening, and belongs to the technical field of biological medicine analysis. Amino and vinyl sulfuryl are covalently modified on the surface of the microarray pore plate to introduce an active site which can be specifically covalently bound with a membrane protein or a tag group, so that directional fixation of a target membrane protein on the surface of the microarray pore plate is realized, and the functionalized microarray pore plate is obtained. A functional microarray pore plate and a specific fluorescent probe are combined to screen a membrane protein ligand, the specific fluorescent probe is combined with a receptor active site of a fixed protein, and a detectable fluorescence signal change is generated by utilizing a competitive replacement mechanism of the probe and a candidate compound at the active site; the method is used for in-vitro screening and discovery of potential membrane protein ligands.

This invention relates to a novel, highly efficient bactericide and its preparation method, belonging to the field of antibacterial materials technology. The invention utilizes the reaction of epoxy groups and tertiary amine compounds in maleic pine carboxylic acid glyceride under hydrochloric acid to convert the epoxy groups into a hydroxyl quaternary ammonium salt structure. Then, an esterification reaction between trimellitic anhydride acyl chloride and hydroxyl groups is used to introduce an anhydride group into the molecular chain. Finally, an esterification reaction between anhydride and hydroxyl groups is used to introduce quaternary phosphonium salt and silver citrate structures into the molecular chain, resulting in a novel, highly efficient bactericide. The novel, highly efficient bactericide prepared by this invention simultaneously contains quaternary ammonium salt, quaternary phosphonium salt, and metallic silver ions. The positively charged quaternary ammonium salt and quaternary phosphonium salt are adsorbed onto the negatively charged bacterial surface through electrostatic interactions, exerting antibacterial and bacteriostatic effects by altering membrane permeability. The silver ions in silver citrate can bind to the sulfhydryl and carboxyl groups of bacterial membrane proteins, disrupting membrane integrity and thus exerting antibacterial and bacteriostatic effects.

The invention provides a detection method for reducing the off-target risk of a targeting drug, and relates to the technical field of immunodetection. The detection method comprises the following steps: screening the optimum working concentration of a test substance, screening the human plasma membrane protein having a binding risk with the test substance, and screening out a target protein having a specific binding relationship with the test substance; according to the detection method provided by the invention, the expression of the fluorescent protein can indicate the expression condition of the human plasma membrane protein, positive control, quality control control and negative control are set, and the target protein having a specific binding relationship with a test substance is obtained through chemiluminescence detection and flow cytometry screening, so that the risk of a false negative result is reduced, and the detection accuracy is improved. And the accuracy of predicting the off-target risk of the targeted drug is improved.