105 results about "Membrane binding" patented technology

Compositions of retroviruses and methods of using the same for gene delivery, wherein the retroviruses comprise a viral envelope protein comprising at least one mutation that diminishes its native function, a non-viral membrane-bound protein comprising a membrane-bound domain and an extracellular targeting domain.

The present invention relates to recombinant binding proteins comprising an ankyrin repeat domain with binding specificity for soluble mesothelin or membrane-bound mesothelin. In addition, the invention relates to nucleic acids encoding such recombinant binding proteins, pharmaceutical compositions comprising such proteins or nucleic acids, and the use of such binding proteins, nucleic acids or pharmaceutical compositions in methods for treating or diagnosing diseases, such as cancer in a mammal, including a human.

The invention discloses a methotrexate-cationic polypeptide conjugate as well as a preparation method and application thereof, and belongs to the technical field of medicinal chemistry. According to the conjugate, MTX and cationic polypeptide with the amino acid sequence shown as SEQ ID NO: 1 are connected through chemical bonding, the membrane binding characteristic of oligomeric lysine and the transmembrane capacity of TAT cell-penetrating peptide are creatively fused, and a polypeptide carrier system with efficient cell penetrating capacity is constructed; mTX obtains amphipathy and electropositivity through peptide fragment modification, stable nanoparticles can be spontaneously formed, and the problems of poor water solubility and instable acidity of raw material medicines are effectively improved; according to the conjugate, the accumulation efficiency of the medicine at a diseased region is remarkably improved by utilizing the targeting characteristic of the polypeptide carrier, the action time of the medicine is prolonged through a precise delivery mechanism, and meanwhile, the system toxicity is reduced. The preparation method adopts mild coupling reaction conditions, and the product is high in purity and excellent in stability; the compound is suitable for treating immune-mediated inflammatory diseases and specific malignant tumors, and provides an innovative solution thought for treating related diseases.

The invention relates to the technical field of gas separation membrane materials, in particular to a preparation method of a PIL-MOF composite membrane and application of the PIL-MOF composite membrane in CO2 / N2 separation.The preparation method comprises the steps that S1, a PET base membrane is pretreated, specifically, the PET membrane is soaked in a 3M HNO3 solution for 8-12 hours, washed with deionized water to be neutral and then dried; s2, in-situ growth of UiO-66-NH2: dissolving zirconium tetrachloride in acetic acid, preserving heat at 80 DEG C for 0.5-1.5 hours, cooling, adding DMF and 2-aminoterephthalic acid to form a precursor solution, dipping the pretreated PET film for 1-3 hours, and performing solvothermal reaction at 110-130 DEG C for 18-24 hours to obtain a PET / UiO-66-NH2 film; and S3, photo-initiation polymerization: coating the surface of the PET / UiO-66-NH2 film with a solution containing [VBIM] [BF4], a cross-linking agent and a photoinitiator, carrying out 365 nm ultraviolet light curing for 0.5-2 h, and drying to obtain the PIL-MOF composite film. The composite membrane combines the porous characteristic of the metal organic framework and the functional characteristic of the ionic liquid, and has application value in the fields of gas separation, catalysis and the like; the preparation process is simple, the interface bonding strength is high, and the product performance is stable.

The invention relates to germline-targeting designs, stabilization designs, and / or combinations thereof, of proteins designed with modified surfaces helpful for immunization regimens, other protein modifications and / or development of nanoparticles, methods of making and using the same, and to (a) germline-targeting priming or boosting / shepherding immunogens to initiate or guide maturation of VRC01-class responses (b) PCT64 / PG9-germline-targeting designs (c) BG18-germline-targeting designs or boosting / shepherding immunogens to initiate or guide maturation of BG18-like responses, and / or (d) trimer stabilization and presentation in a membrane-bound format.

A method for generating or expanding a population of natural killer cells (NK cells) via using modified K562 feeder cells having expression of factors capable of increasing activation and proliferation of natural killer cells, wherein the factors capable of increasing activation and proliferation of NK cells include: (A) a co-stimulatory molecule comprises CD80 molecules, and 4-1BBL molecules; and (B) a cytokine, specifically membrane-bound IL-21 (mbIL-21).

The invention provides a tumor infiltrating lymphocyte expressing a membrane binding cytokine, and particularly provides a modified TIL, and an expression membrane of the modified TIL anchors IL-7. The TIL can be effectively activated and proliferated, can be efficiently detected and sorted, and can effectively mediate a molecular braking effect.

The invention belongs to the field of biological medicine, and discloses a membrane binding type IL7 fusion protein, an engineered immune cell for expressing the membrane binding type IL7 fusion protein and application of the membrane binding type IL7 fusion protein. The fusion protein comprises an IL7 region and a transmembrane domain and can be expressed on a cell membrane, the tumor cell killing ability and the T cell survival ability of T cells expressing the fusion protein are both enhanced, and the aims of improving the tumor immune cell treatment effect and reducing the toxic and side effects are achieved. Particularly, the IL7 fusion protein anchors and expresses IL7 on the surface of a cell through transmembrane domains such as CD80 or PD-L1 and the like, so that (1) immune cells can be accurately regulated and controlled, the possibility that an excessive IL7 signal possibly causes autoimmune response or aggravates CRS is reduced, and the safety of the IL7 to immune cells such as T cells and the like is enhanced; (2) the half-life period of IL7 is prolonged, and the anti-tumor effect of adoptive immune cells is enhanced; and (3) the transmembrane fragment is linked with the IL7 through a hinge region of the flexible linker G4S, CD80 or PD-L1, so that the flexibility of the IL7 is enhanced, and the proliferation and killing functions of the IL7 on T cells are enhanced.

Disclosed herein are methods for treating a disease or disorder of the central or peripheral nervous system by administering to a subject in thereof an agent capable of modulating the activity or expression of bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI). Additionally, methods for screening agents capable of modulating the activity or expression of BAMBI are disclosed.

Synthetic nucleic acids encoding mini and microdystrophin genes comprising the membrane binding motifs or domains of the R10-R11-R12 region are provided. Also provided are vectors, host cells, and related methods of using the same to treat a subject suffering from Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) or X-linked dilated cardiomyopathy (XLDC), or for ameliorating one or more adverse effects of DMD, BMD, or XLDC. Also provided are a fusion protein comprising a nNOS binding domain of dystrophin R16-R17 that is operably linked to a syntrophin PDZ domain and synthetic nucleic acids comprising the same that can be used to treat subjects with diseases characterized by loss of sarcolemmal neuronal nitric oxide synthase (nNOS) activity.

The invention provides a recombinant cordyceps militaris strain with high yield of beta-glucan as well as a construction method and application of the recombinant cordyceps militaris strain, and belongs to the technical field of bioengineering and genetic breeding of edible fungi. According to the invention, a cordyceps militaris CmGls super-safe overexpression vector is constructed through a CRISPR-Cas9 technology, and then the cordyceps militaris CmGls super-safe overexpression vector is transferred into cordyceps militaris to obtain a recombinant cordyceps militaris strain with high yield of beta-glucan. According to the recombinant cordyceps militaris strain, membrane binding type beta-1, 3-glucan synthetase gene CmGls is accurately inserted into a safe port site CmSh1 of a cordyceps militaris genome, compared with a wild type cordyceps militaris strain, the growth rate of the recombinant cordyceps militaris strain is remarkably increased by 15% or above, and yellow phenotype and cell wall stress resistance are remarkably enhanced; after 6 days of submerged fermentation, the mycelium amount and the extracellular beta-glucan yield are respectively improved by more than 10%, and the strain has good application in improving beta-glucan synthesis.

The present invention discloses the construction and application of a recombinant Gluconobacter oxydans, belonging to the fields of genetic engineering and whole-cell catalysis. The present invention heterologously expresses an epoxide hydrolase from Sphingomonas in Gluconobacter oxydans, and simultaneously utilizes the alcohol and aldehyde dehydrogenases bound to the membrane of Gluconobacter oxydans itself to synthesize R-mandelic acid in a one-step process using styrene oxide as a substrate. The present invention combines the transcriptome data of Gluconobacter oxydans, screens 7 strong promoters using green fluorescent protein as a reporter gene, and determines the strongest promoter P for expressing the epoxide hydrolase gene. 12780 , ultimately significantly increasing the yield of R-mandelic acid.

To provide assays for the detection and / or quantification of membrane-associated proteins, e.g., circulating CD20 (cCD20), incorporating an extracellular vesicle-based calibrator comprising the membrane-associated tumor antigen as well as the use of such assays in the detection and treatment of hyperproliferative disorders.SOLUTION: An assay for detecting a membrane-associated protein in a sample, comprises: a) a capture antibody that binds to an extracellular vesicle comprising the membrane-associated protein in the sample, thereby generating a capture antibody-extracellular vesicle complex; and b) a detection antibody that binds to the capture antibody-extracellular vesicle complex to form a detectable bound complex, where a signal from the detectable bound complex is calibrated against one or more known values detected from the extracellular vesicle comprising the protein.SELECTED DRAWING: Figure 1

Provided is a tumor infiltrating lymphocyte expressing a membrane-bound cytokine, specifically a modified TIL expressing membrane-anchored IL-7. The TIL can be effectively activated and proliferated, can be efficiently detected and sorted, and can effectively mediate molecular braking.

Described herein is an engineered viral subunit vaccine encoding ASFV capsid proteins (e.g., P72 and Penton) for use in pigs. The vaccine with either P72 or Penton tested in mice elicited significantly higher levels of antigen-specific IgM and IgG, increased frequency of antibody-secreting cells in the bone marrow, and higher quality germinal centers in the spleens of immunized mice. Enhanced T cell responses were also observed, with higher levels of IFN-γ and TNF-α in splenocytes. Immunogenicity assessments in pigs further supported these findings, with both MB-P72 and MB-PN180Q inducing robust B cell and T cell responses. These findings show that expression of capsid proteins P72 and Penton in membrane-bound forms via mRNA preserves their native multimeric structure and enhances their immunogenicity and provides a basis for an effective ASFV vaccine.

The application belongs to the technical field of biological medicine, and provides a CXCR4 high expression type iPSC-NK cell with enhanced bone marrow and tumor tissue homing ability, and a preparation method and application thereof. The cell takes pluripotent stem cells as starting cells, overexpresses a membrane-bound IL-15 and IL-15RA fusion protein gene and a CXCR4 receptor gene in the pluripotent stem cells; pluripotent stem cells stably expressing the target gene are obtained, and then iPSC-NK cells are obtained through induction differentiation. The membrane-bound IL-15 and IL-15RA fusion protein gene and the CXCR4 receptor gene are targetedly integrated into a safe harbor site of the induced pluripotent stem cell through a gene editing technology, so as to construct an iPSC cell strain stably expressing key proteins; the function-enhanced iNK cell is obtained through induction differentiation, and exhibits excellent bone marrow and various solid tumor tissue homing ability and persistent immune killing activity.

The present invention provides anti-B7H3 antibodies and their application thereof A group of murine anti-B7H3 monoclonal antibodies are prepared by hybridoma technology, using recombinant B7H3 protein as immunogen. Human-murine chimeric antibodies generated from the aforementioned murine anti-B7H3 hybridomas are capable of specifically binding to recombinant B7H3 protein and membrane-bound B7H3 on tumor cell lines. The derived B7H3 scFv-CD3ε-engineered T cells exhibit a significant cytotoxic effect on B7H3-positive tumor cell lines. Therefore, the anti-B7H3 antibodies and B7H3 scFv-CD3ε-engineered T cells have great application prospects in treating or ameliorating B7H3-associated diseases.

The application relates to the technical field of biological medicine, in particular to a genetically modified immune cell and construction and application thereof. The genetically modified immune cell expresses a target protein, the target protein comprises a chimeric antigen receptor and a membrane-bound interleukin, the chimeric antigen receptor comprises an antigen binding domain specifically combined with GPC3, and the membrane-bound interleukin comprises a membrane-bound IL-7. The application simultaneously overexpresses the GPC3CAR and the membrane-bound IL-7, improves the killing ability of TILs on tumor cells, simultaneously improves the expansion ability of immune cells in vivo and in vitro, the ability of continuously killing tumors, and is more conducive to improving the treatment effect of immune cells on GPC3-positive tumors.

Provided are an engineered TIL cell having an improved tumor recognition and killing capability and a use thereof, relating to the technical field of genetic engineering and cell therapy. The surface of the engineered TIL cell comprises an exogenous membrane-bound DAP10-CD3ζ fusion protein, and the endoplasmic reticulum or Golgi apparatus of the engineered TIL cell comprises a retained fusion protein. Specifically, provided is the membrane-bound DAP10-CD3ζ fusion protein, so that the TIL cell can be activated by an activated fusion protein when recognizing a tumor cell by means of the NKG2D-NKG2DL pathway, so as to effectively kill the tumor cell. Further provided is the retained fusion protein, so that endogenous NKG2DL that is expressed when the TIL cell is activated can bind to the retained fusion protein so as to be retained in the cell, thereby avoiding mutual recognition and killing between the TIL cells and improving the viability of the TIL cells.

Protein and RNA-based condensates are emerging as an alternative to classical membrane-bound organelles for the task of compartmentalizing molecules and biochemical reactions. We describe methods for making RNA condensates in mammalian cells. We further show that aptamers make it possible to recruit peptides and proteins to the condensates with high specificity. Such RNA condensates can be modularly customized and offer a route toward creating systems of functional artificial organelles.

Provided are genetically engineered induced pluripotent stem cells (iPSCs) and derivative cells thereof expressing a chimeric antigen receptor (CAR) and a membrane bound IL-12 and methods of making and using the same. Also provided are compositions, polypeptides, vectors, and methods of manufacturing.

This invention discloses a multifunctional engineered NK cell, its construction method, and its applications. This NK cell co-expresses a membrane-bound targeted cytokine complex and a secretory bispecific nanobody via a single-carrier system. The membrane-bound complex uses an anti-PD-L1 single-domain antibody to directionally anchor IL-15 / IL-21 to the cell membrane surface and utilizes synaptic recruitment to achieve high-level enrichment of cytokine signals at the immune synapse, enabling precise paracrine secretion of cytokine signals. The secretory bispecific antibody mediates specific cytotoxicity by transdirectionally linking the NK cell activation receptor NKp46 with the tumor-associated antigen B7-H3. Experiments have demonstrated that the engineered NK cells constructed in this invention significantly enhance the killing efficacy against PD-L1 or B7-H3 positive tumor cells while maintaining a high P2A cleavage rate, and the synergistic index shows a significant synergistic effect. This invention effectively overcomes the technical shortcomings of traditional NK cell therapy, such as systemic cytokine toxicity and tumor antigen escape, providing a novel strategy for immunotherapy of solid tumors.

Methods and a recombinant C1- fixing microorganism for the continuous production of products from gaseous substrates. Further, the gaseous substrate comprises CO2 and an energy source. The recombinant C1-fxing microorganism has a disruptive mutation in a membrane bound hydrogenase gene. The C1-fixing microorganism having the disruptive mutation minimizes the H2:CO2 uptake ratio.

The application discloses a kind of based on HIV envelope protein chimeric exosome and its preparation method and application, belong to biological medicine technical field.The application first constructs the cell line of stable expression HIV envelope protein Env, obtains engineered exosome from cell culture supernatant separation and purification;The exosome is used as immunogen combined with adjuvant immunization experimental animal, and high-efficiency induction specific humoral immune response;Again, obtain Env antigen specificity single B cell by flow cytometry sorting, obtain antibody variable region gene by single cell lysis, reverse transcription and nest PCR amplification, cloning to expression vector and expressing in mammalian cell, finally, HIV specific neutralizing antibody is screened by binding activity, affinity and neutralizing activity;The application is combined with single B cell antibody screening technology by embedding HIV Env antigen in the form of membrane combination in exosome surface, and realizes the synergistic optimization of antigen delivery and antibody screening process.

Disclosed is a method for preparing enantiomerically enriched compound 1, which involves chiral salt resolution of compound 1, or intermediate INT 1f or intermediate INT 1g. Compound 1 is useful for treating diseases or conditions, such as cancer, particularly diseases or conditions dependent on the activity of membrane-bound tyrosine- and threonine-specific cdc2 inhibitory kinase (Myt1). JPEG2025542335000078.jpg43170

The present invention is in the fields of antibody technology, medicine, pharmacology, infection biology, and medical diagnostics. More specifically, the present disclosure provides VHH antibodies that prevent membrane binding and / or oligomerization of Staphylococcus aureus (HLA) alpha-hemolysin and HLA-mediated hemolysis.

The invention relates to a vaccine platform, comprising a lipid binding amino acid sequence and an oligomerization sequence. In particular, the lipid binding amino acid sequence and an oligomerization sequence are derived from filensin, a protein with no or minimal immunogenicity. Filensin has an extremely strong membrane binding capacity and oligomerization property, making it an ideal carrier for an antigenic moiety. An immunization platform comprising a nucleic acid sequence(s) coding for a lipid binding amino acid sequence and an oligomerization sequence is also provided.

The invention discloses a yeast immunochromatography rapid detection test strip and a preparation method thereof, and relates to the technical field of immunodetection.The test strip comprises a functionalized sample pad, a combination pad, a reaction film and a water absorption pad which are in lap joint in sequence; the sample pad is pretreated by a treating fluid containing beta-1, 3-glucanase, casein and TritonX-100, so that in-situ wall breaking and interference resistance can be realized; the reaction film detection line is coated with mannose binding lectin and a saccharomycetes specific monoclonal antibody, the binding pad fixes different epitope detection antibodies labeled by colloidal gold, the preparation method comprises the steps of functionalizing the sample pad, coating the reaction film, preparing the binding pad and assembling the test strip, and the matched kit comprises the test strip and a phosphate buffer diluent with a specific pH value. The method can realize extraction-free one-step rapid detection of saccharomycetes, has both specificity and stability, and is suitable for multi-scene screening.

The presently disclosed subject matter provides compositions and systems for cell-based immunotherapy. In certain non-limiting embodiments, the system comprises a membrane-bound polypeptide and at least one soluble polypeptide that is capable of dimerizing with the membrane-bound polypeptide.

Provided herein are engineered T-celi receptors (TCRs) having nanomoiar affinity for the immuno-dominant pp65 peptide residing between residues 495-503 (NLV) in complex with HLA-A2*02:01. The TCRs may be membrane-hound TCRs, soluble TCRs, chimeric TCRs, or chimeric antigen receptors. Also provided are methods of using the engineered TCRs to treat diseases, monitor disease progression, monitor vaccine efficacy, and detecting NLV / A2 presentation on the surface of cells.