53 results about "Membrane binding" patented technology

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.

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.

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.

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.

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.

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.

The invention discloses a CAR-T cell modified by an interleukin 15 receptor alpha mutant and application of the CAR-T cell, and particularly relates to a gene engineering medicine. The CAR-T cell is integrated with a multi-gene expression unit through an SB100X transposon system, wherein the multi-gene expression unit comprises opti-mIL-15 (T62G / L55F), opti-mIL-15R alpha-Sushi (Q199E / D203A), a GPI anchoring sequence, a (G4S) 3 flexible connecting peptide, a targeted CAR gene and an iC9 safety switch, and a membrane binding type IL-15 / IL-15R alpha compound supported by an autocrine signal is formed. The affinity and stability of IL-15 signals are enhanced through mutants, efficient integration and co-expression are achieved through non-viral vectors, the in-vivo durability, amplification capacity and anti-tumor activity of CAR-T cells are remarkably improved, and meanwhile a cell factor local delivery and induction type safety switch system is combined through a membrane.

The invention discloses a targeted receptor ubiquitination induced antibody fusion protein coupling drug and application thereof, and the antibody fusion protein coupling drug comprises: (a) an antibody fusion protein, which comprises an antibody structural domain capable of binding to a tumor-associated antigen receptor; the membrane binding type E3 ubiquitin ligase binding module is fused with the antibody structural domain, and the E3 ubiquitin ligase binding module is an RSPO2 FU (F109A) structural domain; wherein the antibody fusion protein can be simultaneously combined with a tumor associated antigen receptor and a membrane binding type E3 ubiquitin ligase RNF43 and / or ZNRF3; and (b) a cytotoxic small molecule drug coupled with the antibody fusion protein. According to the invention, by introducing a membrane binding type E3 ubiquitin ligase binding module, the antibody can promote ubiquitination modification and lysosomal pathway degradation while binding to a tumor-associated antigen receptor, so that the internalization efficiency and tumor killing activity of the antibody-conjugated drug are significantly improved.

The methods and compositions disclosed herein relate to the field of cell therapy, and more specifically, to improving CAR and / or TCR function through improvement of the tumor microenvironment via improvement in cytokine signaling. Methods of treating a cancer in a patient are disclosed comprising administering to the patient immune cells expressing a chimeric antigen receptor or T-cell receptor and a membrane bound IL-18. Further disclosed are nucleic acids which encode a membrane bound IL-18 and methods for culturing cells expressing such nucleic acids.

The present application relates to the technical field of biomedicine, and in particular to a genetically modified immune cell, and construction and use 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 that specifically binds to GPC3, and the membrane-bound interleukin comprises a membrane-bound IL-7.

The present disclosure provides methods and compositions for genetically modifying lymphocytes and related methods that include genetically modifying T cells and / or NK cells. The methods use replication incompetent recombinant retroviral particles that comprise a pseudotyping element on their surface and optionally a membrane-bound T cell activation element, such as an anti-CD3, and encode one or more engineered signaling polypeptides that can include a lymphoproliferative element, and / or a chimeric antigen receptor (CAR). The methods can include contacting PBMCs with replication incompetent recombinant retroviral particles for various exemplary time periods, such as less than 24 hours or in some illustrative embodiments less than 15 minutes. In some aspects, the present disclosure provides methods and compositions for genetically modifying lymphocytes, for example T cells and / or NK cells, in whole blood or a component thereof. In some embodiments a lymphodepletion filter assembly is used before or after forming a reaction mixture where lymphocytes are contacted with recombinant retroviral particles in a closed system, to genetically modify the lymphocytes.

Disclosed herein are 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.

Provided is a dual-target BCMA / CD19 chimeric antigen receptor construct. Membrane-bound interleukin-15 is fused, via a self-cleaving peptide, at the C-terminus of a bispecific CAR targeting BCMA and CD19, and the CAR construct is introduced into a DNT cell, such that a universal dual-target BCMA / CD19-CAR-DNT cell having enhanced function is obtained. The cell simultaneously targets BCMA and CD19, and has cell-killing activity, thereby providing a new treatment for BCMA- and / or CD19-mediated diseases.

The invention provides an NK cell, a chimeric antigen receptor and a pharmaceutical composition and application of the chimeric antigen receptor, the NK cell expresses the chimeric antigen receptor, and the chimeric antigen receptor fusion protein comprises a targeted TROP2 single-chain antibody, a transmembrane structural domain, a co-stimulation structural domain, an activation structural domain, IL-15 protein and a CXCR2 chemotactic factor from the N terminal to the C terminal. In the invention, overexpression of CXCR2 can improve the killing efficiency of CAR NK, and the killing efficiency can be improved only by combining the membrane-combined IL15 with CXCR2 in the two forms of IL15.

The disclosure provides, in various embodiments, polynucleotides encoding a fusion protein, wherein the polynucleotides comprise: (a) a protein coding sequence encoding a non-Hedgehog therapeutic protein, and (b) at least one of the following: (i) a sequence encoding a polypeptide sufficient for palmitoylation by a Hedgehog acyltransferase, wherein the polypeptide is N-terminal to the non-Hedgehog therapeutic protein, (ii) a sequence encoding a Hedgehog auto-processing domain having sterol modification activity, wherein the domain is C-terminal to the non-Hedgehog therapeutic protein, and (iii) a sequence encoding a polypeptide sufficient for palmitoylation by a membrane-bound O-acyltransferase enzyme. The disclosure also provides, in various embodiments, vectors, cells, and kits comprising the polynucleotides, and methods of using the polynucleotides, vectors, cells, and / or kits.

A synthetic cytokine signaling system comprises an engineered cytokine signaling chain encoding a transmembrane polypeptide containing a specific variant intracellular domain of IL9R and an extracellular domain of cytokine receptors such as IL2R [beta], IL4R [alpha], IL7R [alpha], IL9R and IL21R. The engineered cytokine signal chain may be in a self-activating form, or optionally may be in a non-self-activating form, depending on whether the functionally compatible cytokine ligand domain is fused to the N-terminus. In the latter case, the synthetic cytokine signaling system may further comprise an engineered cytokine ligand chain that encodes a corresponding cytokine ligand, which may be membrane-bound or secreted. Immune cells modified with the synthetic cytokine signaling system exhibit improved properties compared to unmodified immune cells, such as zero or reduced dependency on IL-2 when cultured in vitro, enhanced activation after antigen stimulation, enhanced cytotoxicity to target cells, and improved tumor control ability when metastatic in vivo.

To provide methods for degrading target surface proteins in the ubiquitin pathway using bispecific binding agents or immunoconjugates that bind to the target surface proteins and membrane-bound ubiquitin E3 ligase.SOLUTION: The present invention relates, inter alia, to methods of degrading target surface proteins in the ubiquitin pathway using bispecific binding agents or immunoconjugates that bind to target surface proteins and membrane-bound ubiquitin E3 ligase. The present invention also relates to methods of degrading target surface proteins in the ubiquitin pathway using engineered membrane spanning proteins that bind to target surface proteins and exhibit ubiquitin E3 ligase activity. The present disclosure also provides compositions and methods useful for making bispecific binding agents and engineered transmembrane proteins, immunoconjugates, nucleic acids encoding them, host cells genetically modified with said nucleic acids, and methods of modulating the activity of cells and / or treating various diseases, such as cancer.SELECTED DRAWING: Figure 1

To provide an antigen-binding protein which binds to a membrane-bound target and can be internalized. Also provided is an immunoconjugate comprising the antigen binding protein and a cytotoxic agent.SOLUTION: Provided is an antigen binding protein that specifically binds to BCMA and inhibits the binding of BAFF and / or APRIL to BCMA, wherein the antigen binding protein is capable of binding to Fc γ RIIIA or is capable of Fc γ RIIIA-mediated effector function and is capable of internalization.SELECTED DRAWING: Figure 1

There is provided a composition for modifying a T cell, the composition comprising: a protein complex comprising a polynucleotide-modifying enzyme domain, a T cell membrane binding domain and an endosome escape domain; a guide oligonucleotide specific to a T cell receptor constant (TRAC) gene of the T cell; and a donor DNA comprising two homology arms at each end of the donor DNA homologous to exon1 of the TRAC gene and encoding therebetween a chimeric antigen T cell receptor comprising: translocation signal for translocation to a cell membrane of the T cell; a transmembrane domain; an intracellular signaling domain; and an extracellular antigen binding domain.