129results about "Polypeptide with Strep-tag" patented technology

The invention specifically discloses a composition for detecting NAD < + > and application thereof, and relates to the technical field of biological detection. According to the composition for detecting NAD < + >, the composition comprises an NAD < + > binding protein fragment A connected with an Fc tag, an NAD < + > binding protein fragment B and a connecting peptide, the detection result is accurate and reliable, and meanwhile the technical difficulty, uncertainty and high cost faced by antibody screening in a traditional method are avoided.

The present invention relates to a fusion protein that produces point mutations in cells, its preparation and use. Specifically, the fusion protein provided by the present invention contains a Cas enzyme that lacks cytosine deaminase and nuclease activity and retains helicase activity, or is formed by a Cas enzyme that lacks cytosine deaminase and nuclease activity and retains helicase activity. The present invention also relates to the coding sequence of the fusion protein, a polynucleotide sequence containing the coding sequence, a nucleic acid construct containing the polynucleotide sequence, a corresponding host cell, a method for producing point mutations in a cell, and a kit, etc. By using the present invention, it is possible to achieve site-directed mutagenesis while obtaining high mutation efficiency and a variety of mutation combinations in a specific gene region.

The invention relates to mutant forms of CsgG. The invention also related to analyte detection and characterisation using CsgG.

The present invention relates to modified Dda helicases which can be used to control the movement of analytes such as polynucleotides. The modified Dda helicases are used in analyte detection and characterisation. The present invention also relates to novel protein pores and their uses in analyte detection and characterisation. The invention particularly relates to an isolated pore complex formed by a CsgG-like pore and a modified CsgF peptide, or a homologue or mutant thereof, thereby incorporating an additional channel constriction or reader head in the nanopore.

Provided herein are methods for preparing and characterizing SARS-co2 antigen specific immune cell cultures and preparations and methods of using the same in adoptive immunotherapy for cancer, infections and immune disorders. Also, provided are compositions and methods for generating immune cells expressing synthetic antigen binding receptors targeting SARS-cov2 and methods of use of these cells for the treatment and prevention of COVID-19. Also provided are compositions and methods for determining immune response to SARs-cov2 in a subject, detecting SARS-cov2, measuring cytotoxicity induced by SARS-cov2, and detecting the expression and cytotoxicity of synthetic antigen binding receptors targeting SARS-cov2.

The invention provides preparation and application of an improved recombinant III-type collagen peptide. The collagen comprises an amino acid sequence as shown in SEQ ID NO.1, and a tag sequence is fused at the N terminal and / or C terminal of the amino acid sequence as shown in SEQ ID NO.1. The recombinant collagen provided by the invention only contains a small affinity tag during expression, is convenient to purify and does not need enzyme digestion, so that the process steps are greatly simplified, the process cost is reduced, the excellent characteristics of the original recombinant collagen peptide Co8 (SEQ ID NO.1) are maintained, and the recombinant collagen still has high expression quantity, high hydrophilicity and high stability.

Provided is a fusion protein, comprising a first binding domain that specifically binds a tumor-associated antigen, and a second binding domain that specifically binds a CD47 protein, wherein the second binding domain comprises a mutant of a human SIRPα variant 1. Also provided are an immunoconjugate, nucleic acid molecules, vectors, compositions, and cells related to the fusion protein and preparation methods therefor, and the uses thereof in the preparation of medicaments.

The present invention involves an engineered probiotic bacterium comprising a heterologous nucleic acid, where the heterologous nucleic acid comprises a nucleic acid sequence encoding an anti-spike glycoprotein nanobody of a coronavirus. In one embodiment, the bacterium is Escherichia coli Nissle 1917. In another embodiment, the anti-spike glycoprotein nanobody appears on the surface of the probiotic bacteria.

To provide a system including novel nanopores having different structures and recognition sites that improve sequencing accuracy and / or provide different error profiles.SOLUTION: Provided are systems relating to the field of nanopores and their use in various applications, e.g., analysis of biopolymers and macromolecules, typically by making electrical measurements during translocation through the nanopore, comprising a funnel-shaped proteinaceous nanopore comprising alpha-helical pore-forming toxins that are members from the actinoporin family of proteins, more particularly Fraga Ceatoxin C (FraC), a variant FraC, a FraC paralog or a FraC homolog.SELECTED DRAWING: None

Provided are a single-chain polypeptide which is highly activated after entering a specific use site, attenuated and can be safer and more convenient to produce, and the corresponding use thereof, a nucleic acid encoding same and a preparation method therefor. The single-chain polypeptide comprises: a first domain, an intermediate amino acid sequence region, and a second domain, wherein the intermediate amino acid sequence region is located between the first domain and the second domain, and contains at least one first enzyme cleavage site, the first enzyme cleavage site is designed to be capable of being highly specifically cleaved by a first protease present in a specific part of the human body, and the single-chain polypeptide is converted into an activated form capable of inhibiting the release of an information substance after being cleaved by the first protease at the first enzyme cleavage site.

The present invention provides a combination comprising a) an antigen binding domain specific for CD90, and b) an antigen binding domain specific for CD326, for use in treatment of human cancer comprising cancerous cells that co-express CD90 and CD326. In one embodiment of the invention the combination comprises a) an immune cell comprising a CAR comprising an antigen binding domain specific for a tag of a first and a second polypeptide, b) said tagged first polypeptide that has an antigen binding domain specific for CD90, and c) said tagged second polypeptide that has an antigen binding domain specific for CD326, wherein the tag of the first polypeptide and the tag of the second polypeptide are identical. In a further embodiment the concentrations used for said first and that second polypeptide are below the activation threshold of said CAR, respectively, but the sum of both concentrations is above the activation threshold of said CAR.

The invention relates to a method for identifying and quantifying a polypeptide from a library of polypeptides. The method comprises the steps of: 1 - providing a polypeptide library and a detection tag library, 2 - generating a nested library comprising the polypeptides and the detection tags, 3 - sequencing the nested library, 4 - selecting a member of the nested library in one or several selection steps that are independent of a physical genotype-phenotype linkage, 5 - isolating the detection tag from the selected polypeptide, 6 - identifying and quantifying the detection tag by mass spectrometry, 7 - obtaining the sequence of the selected polypeptide.

Herein is reported a method for producing C-terminally biotinylated FcRn comprising the steps of cultivating a mammalian cell comprising a deoxyribonucleic acid encoding FcRn and E. coli biotin-[acetyl-CoA-carboxylase] ligase (BirA) in a biotin containing medium, and recovering C-terminally biotinylated FcRn from the cell or the cultivation medium, wherein the deoxyribonucleic acid encoding FcRn and E. coli BirA is stably integrated into the genome of the mammalian cell and comprises in 5′- to 3′-direction a first expression cassette encoding class I major histocompatibility complex-like protein (α-FcRn) comprising a HisAvi-tag at the C-terminus, a second expression cassette encoding β2-microglobulin (β2m), a third expression cassette encoding class I major histocompatibility complex-like protein (α-FcRn) comprising a HisAvi-tag at the C-terminus, a fourth expression cassette encoding β2-microglobulin (β2m), and a fifth expression cassette encoding E. coli biotin-[acetyl-CoA-carboxylase] ligase.

The invention discloses a chimeric antigen receptor exosome targeting a CD8T cell, a CAR-T cell and a construction method and application of the chimeric antigen receptor exosome, and relates to the technical field of biomedicine, and the construction method comprises the following steps: co-transfecting three plasmids, namely BRD-PTK-CD8 < + >-Lamp2b, pcDNA3.1-CD63-L7Ae and pcDNA3.1-N6 (SP)-Nb60-T20-C / D box, into a 293F cell, incubating, and purifying to obtain the exosome capable of targeting the CD8T cell. The exosome can deliver CAR-mRNA to T cells and convert CD8 + cells into CAR-T cells, and blood of an infected person does not need to be drawn; cAR molecules aiming at in-vivo malignant cells can be prepared in vivo; chemotherapy and radiotherapy are not needed, and side effects are avoided; and universality is achieved.

To provide compositions and methods to improve delivery of gene therapy vectors and therapeutics across the BBB to provide more effective treatment for a variety of CNS conditions.SOLUTION: Compositions and methods for delivering effector entities to the CNS of a subject are provided. Engineered AAV capsids that bind GPI-anchored proteins on the BBB are provided as well as methods for their use, including delivery of gene therapy and effector entities. Also provided are methods for reducing the infectivity of the CNS by an AAV.SELECTED DRAWING: None

In the field of biological therapy, a major hurdle is the need for a platform technology that enables high bioavailability and long therapeutic exposure of a biological therapy in the setting of localized disease. Classical approaches result in problematic trade-offs such as toxicity and off-target side effects. The present disclosure provides compounds comprising follistatin domain 1 (FSD1) of follistatin (FST) which are able to bind biological structures, such as the extracellular matrix, through heparan sulfates, without bearing the undesirable neutralizing effects on activin A, myostatin, and GDF11 activities of e.g. full-length follistatin. The invention relates to fusion proteins or conjugated proteins, and compositions thereof, constructs or vectors encoding thereof, medical uses thereof and methods for biological therapy.

The invention discloses an engineered S-layer and a preparation method thereof, an HEMT sensor based on in-vitro self-assembly of the engineered S-layer and a preparation method of the HEMT sensor and a biomolecule detection system, the engineered S-layer is formed by fusion expression of S-layer protein based on recombination or redesign and a biological sensitive element, and the biological sensitive element is protein or polypeptide; the HEMT sensor based on engineering S-layer in-vitro self-assembly comprises a field effect transistor body and a grid sensitive region, the field effect transistor body comprises a source electrode and a drain electrode which are arranged on the two sides respectively. The grid electrode sensitive region is arranged between the source electrode and the drain electrode and comprises a semiconductor material for forming a heterojunction structure and a two-dimensional protein nano array containing a biological sensitive element, and the two-dimensional protein nano array is formed by in-vitro self-assembly of an engineered S-layer on the surface of the semiconductor material. The biomolecule detection system comprises an HEMT sensor based on engineering S-layer in-vitro self-assembly, an electrical detection system and a signal processing and analyzing system.

The invention discloses a method for promoting yarrowia lipolytica to synthesize carotenoid, which comprises the following steps of: recombining and expressing beta-carotene hydroxylase CrtZ and / or beta-carotene ketolase CrtW fused with a short peptide label in the yarrowia lipolytica producing beta-carotene; site-specific mutagenesis is carried out on the amino acid site of CrtZ; and a CrtZ coding gene is integrated in a multi-copy manner. The invention also discloses a yarrowia lipolytica genetically engineered bacterium for producing beta-carotene, wherein carotenoid is selected from zeaxanthin, canthaxanthin and astaxanthin. The yield of zeaxanthin, canthaxanthin and astaxanthin in yarrowia lipolytica is remarkably increased through enzyme label engineering, combinatorial optimization and site-specific mutagenesis strategies, and a new path is provided for efficient synthesis of carotenoids.

This invention relates to fusion polypeptides comprising a DNAX-activating protein 10 (DAP10) polypeptide and a DNAX-activating protein 12 (DAP12) polypeptide. The disclosure also relates to cells comprising such fusion proteins and their use in treating cancer.

Described herein are reagents and methods for high throughput screening of antigens that bind to MHCII alleles.

The present invention relates to modified Dda helicases that can be used to control the movement of analytes, such as polynucleotides. The modified Dda helicases are used in the detection and characterization of analytes. The present invention also relates to novel protein pores and their use in the detection and characterization of analytes. The present invention particularly relates to isolated pore complexes formed by CsgG-like pores and modified CsgF peptides, or homologs or variants thereof, thereby incorporating an additional channel constriction or reader head within the nanopore.

A clustered regularly interspaced short palindromic repeat (CRISPR)-associated complex for adaptive antiviral defence (Cascade); the Cascade protein complex comprising at least CRISPR-associated protein subunits Cas7, Cas5 and Cas6 which includes at least one subunit with an additional amino acid sequence possessing nucleic acid or chromatin modifying, visualising, transcription activating or transcription repressing activity. The Cascade complex with additional activity is combined with an RNA molecule to produce a ribonucleoprotein complex. The RNA molecule is selected to have substantial complementarity to a target sequence. Targeted ribonucleoproteins can be used as genetic engineering tools for precise cutting of nucleic acids in homologous recombination, non-homologous end joining, gene modification, gene integration, mutation repair or for their visualisation, transcriptional activation or repression. A pair of ribonucleotides fused to FokI dimers may be used to generate double-strand breakages in the DNA to facilitate these applications in a sequence-specific manner.

The present invention relates to mutant forms of the outer membrane-located lipoprotein CsgG, in particular modifications at one or more of positions Tyr51, Asn55 and Phe56. The invention also relates to the use of said mutant CsgG for analyte detection and characterization.

Described herein are isolated monovalent antibodies or antibody fragments thereof that bind human properdin. Such antibodies are useful in methods of treatment for diseases mediated by alternative complement pathway dysregulation.

The present invention discloses a method for increasing the yield of European oyster lysozyme, and belongs to the field of bioengineering. The present invention screens and obtains European oyster-derived lysozyme with relatively high enzyme activity from 12 different sources of lysozyme gene sequences from vertebrates, mollusks, poultry and insects, and realizes the active expression of the enzyme in Pichia pastoris. The present invention also fuses short peptides and protein tags of different amino acids to the N-terminus of OElyz. When the Sumo tag is fused, the extracellular enzyme activity of recombinant OElyz reaches 382.6U / mL at the shake flask level, and this yield level can meet the application requirements of lysozyme in the fields of medicine, biotechnology and food.

The present disclosure relates to RSV F protein mutants, nucleic acids or vectors encoding a RSV F protein mutant, compositions comprising a RSV F protein mutant or nucleic acid, and uses of the RSV F protein mutants, nucleic acids or vectors, and compositions.

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.

Disclosed by the present application are an antibody library construction method and an application thereof. The method comprises the following steps: inserting a first element and a second element into a same vector or different vectors, and transfecting the vectors into the cells to obtain an antibody expression cell library, i.e., the antibody library. The first element comprises CIS activators and selection marker genes; the second element comprises extracellular antibody library coding domain, Notch nuclear structure domain and intracellular transcription structure domain.

The invention discloses a method for promoting yarrowia lipolytica to synthesize carotenoid, which comprises the following steps of: recombining and expressing beta-carotene hydroxylase CrtZ and / or beta-carotene ketolase CrtW fused with a short peptide label in the yarrowia lipolytica for producing beta-carotene; wherein the CrtZ and / or CrtW fused with the oligopeptide tags are / is formed by (1) inserting CrtZ added with different oligopeptide tags into a yarrowia lipolytica genome, (2) inserting CrtW added with different oligopeptide tags into the yarrowia lipolytica genome, or (3) inserting CrtZ added with different oligopeptide tags and CrtW added with different oligopeptide tags into the yarrowia lipolytica genome; and the positions of the CrtZ and the CrtW added with the short peptide tags are N ends or C ends. According to the invention, the yield of carotenoids synthesized by yarrowia lipolytica is improved by recombinant expression of beta-carotene hydroxylase CrtZ and / or beta-carotene ketolase CrtW fused with short peptide tags in yarrowia lipolytica.