95 results about "CD16" patented technology

Recombinant NK cells, and especially recombinant NK-92 cells express a chimeric antigen receptor (CAR) having an intracellular domain of FcεRIγ. Notably, CAR constructs with an intracellular domain of FcεRIγ had a substantially prolonged duration of expression and significantly extended cytotoxicity over time. The CAR may be expressed from RNA and DNA, preferably as a tricistronic construct that further encodes CD16 and a cytokine to confer autocrine growth support. Advantageously, such constructs also enable high levels of transfection and expression of the recombinant proteins and provide a convenient selection marker to facilitate rapid production of recombinant NK / NK-92 cells.

The invention discloses a kit and method for detecting leukemia and lymphoma based on full-spectrum flow cytometry, the kit comprises 25 antibodies, the antibodies are specifically bound with fluorescein respectively, and leukemia and lymphoma are detected through full-spectrum flow cytometry; the 25 kinds of antibodies comprise HLA-DR (human leukocyte antigen-DR), CD38, CD7, CD34, Lambda, CD19, CD64, CD14, CD5, CD123, CD16, CD20, Kappa, CD117, CD13, CD45, CD11b, CD2, CD10, CD8, CD15, CD4, CD3, CD56 and CD33. The kit comprehensively covers development stages of various lines of bone marrow cells, and common abnormal expressions of various leukemia, myelodysplastic syndromes and lymphoma, and can preliminarily screen various leukemia and lymphoma.

A recombinant NK-92 cell has a constitutive active promotor that effects expression of native CD16, and most preferably homogenous CD16 158V. Further contemplated recombinant NK-92 cells also include a recombinant nucleic acid that encodes an intracellularly retained interleukin (e.g., IL-2 or er-IL-2), wherein the recombinant NK-92 cell will secrete no more than 5,000 pg / mL IL-2 into a culture medium. The recombinant NK-92 cells presented herein have a significantly improved signal-to-noise ratio and exhibit reduced non-ADCC cytotoxicity.

Provided herein are compositions of NK-92™ cells that express a combination of PD-L1 CAR, CD16 and IL2, and the method of using these cells to to reduce tumor cells and cells in tumor microenvironment (e.g., MDSCs or TAMs) and treat cancer.

Provided herein are compositions of NK-92™ cells that express a combination of PD-L1 CAR, CD16 and IL2, and the method of using these cells to to reduce tumor cells and cells in tumor microenvironment (e.g., MDSCs or TAMs) and treat cancer.

Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells and a culture medium composition thereof. Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells or NK cells and a culture medium composition thereof. The NK cells obtained using the provided culture method have high purity and good in vitro expansion effect; the yield of iNK cells is high, and a single iPSC can differentiate into approximately 2,000 NK cells; and the obtained iNK cells highly express CD16 (over 70%). As a result, the problem of low CD16 expression in iNK cells in the prior art, which requires genetic modification methods to solve, has been addressed.

This invention provides an anti-CD16 antibody, the amino acid sequence of which is selected from SEQ ID NO. 1-3 in the heavy chain CDR region and from SEQ ID NO. 4-6 in the light chain CDR region. This invention belongs to the field of antibodies. It provides a protein sequence of an anti-human CD16 mouse monoclonal antibody, and designs a gene expression base sequence based on the sequenced protein sequence. A stable CHO cell line for recombinant expression of this gene is established. The purified antibody can specifically recognize the human CD16 antigen, and the antibody expression yield is much higher than that of antibodies prepared using traditional hybridoma cells. Furthermore, by replacing mouse IgG1 with rabbit IgG while retaining the variable region, a stable CHO cell line for recombinant expression of the rabbit monoclonal antibody is established. The purified antibody maintains its specific recognition of the human CD16 antigen.

Disclosed herein are monospecific and multispecific single chain antibodies having specificity for one or more of CD47, PD-L1, HSA, CD33, LAG3, and CD16.

The invention relates to the technical field of cell culture, and provides an NK cell in-vitro amplification culture method which comprises the following steps: S1, separating to obtain a mononuclear cell of peripheral blood, resuspending the mononuclear cell by using a serum-free NK cell basic culture medium, and then inoculating the mononuclear cell to an NK cell activation culture medium for incubation; s2, culturing to the third day, supplementing the NK cell to activate the culture medium for amplification, then supplementing the NK cell to activate the culture medium once every other day, and culturing to the 19th day to obtain high-purity NK cells; the NK cell activation culture medium is a serum-free NK cell basal culture medium which contains IL-2 (Interleukin-2), IL-12 (Interleukin-12), IL-18 (Interleukin-18), IL-21 (Interleukin-21), a CD16 antibody, a CD56 antibody, 4-1BBL (4-1BBL), nicotinamide, sodium diethyldithiocarbamate, polyinosinic acid and plasma. The method is simple and convenient to operate and short in culture period, the amplification efficiency reaches up to 1050 times within 19 days, and the NK cells are high in purity and accord with clinical treatment standards.

Disclosed herein are monospecific and multispecific single-chain antibodies specific for one or more of CD47, PD-L1, HSA, CD33, LAG3, and CD16.

This invention discloses a method for preparing NK cells, the NK cells obtained thereby, and their applications. The method includes: sorting NK cells from peripheral blood of an allogeneic donor; culturing them in a serum-free medium containing CD3 monoclonal antibody, CD16 antibody, and IL-2, without the need for a feeder layer; preferably, IL-21 is added for amplification on days 3-5 of culture, and the cells are cultured for 20-40 days. This method yields non-genetically modified NK cells, among which CD56... bright The NK cells comprise >90% of the cell line and highly express a variety of activating and functional molecules. These NK cells effectively clear Aβ oligomers and senescent cells, and alleviate neuroinflammation. In treatment, intracranial injection achieves equivalent or better efficacy with only about 1 / 10th the dose administered intravenously, significantly clearing Aβ deposits in the brains of AD model mice and improving pathology.

Disclosed herein are monospecific and multispecific antibodies specific for CD28 and CD3E, and one or more of OX-40, CD40, 4-1BB, HSA, IL-22, PD-L1, CD16, CD47, CD33, LAG3, and EGFR.

The present invention relates to NK cells expressing dual-targeting chimeric antigen receptors for CD19 and CD22, and uses thereof. In the present invention, it was found that the delivery efficiency of a CAR expression vector and the production efficiency of CAR-NK cells are excellent when, among the cell surface proteins of PBMCs for producing CAR-NK cells (UCI-101), CD16 is expressed at 70% or less, natural killer group 2D (NKG2D) is expressed at less than 10%, CD57 is expressed at 30% or less, low-density lipoprotein receptor (LDLR) is expressed at 0.1% or more, and natural cytotoxicity triggering receptor 3 (NKp30) is expressed at less than 10%. In addition, optimal conditions for inducing NK cell differentiation and optimal conditions for transduction, under which the delivery efficiency of a CAR expression vector and the production efficiency of CAR-NK cells are excellent, were established, and CAR-NK cells produced by the method according to the present invention were found to exhibit an excellent antitumor effect in an animal model, and thus can be effectively used as a composition for preventing or treating diseases related to CD22 (or CD19) expression or diseases related to B cells.

The invention discloses a mouse anti-human CD16 recombinant antibody as well as a preparation method and application thereof, and belongs to the technical field of monoclonal antibody preparation. In a light chain variable region of the mouse anti-human CD16 recombinant antibody, the amino acid sequence of CDR-L1 is RASENIYRYLA, the amino acid sequence of CDR-L2 is NAKTLAE, and the amino acid sequence of CDR-L3 is QHHYRTPYT; the amino acid sequence of the CDR-H1 in the heavy chain variable region is TYWMH, the amino acid sequence of the CDR-H2 in the heavy chain variable region is EINPSNGRTNYNEKFKT, and the amino acid sequence of the CDR-H3 in the heavy chain variable region is NYYGPFPY. According to the preparation method, light and heavy chain variable region sequences are connected through Linker and inserted into an eukaryotic expression vector containing an Fc gene, then CHO-S cells are transfected, the disulfide bond mismatch probability is reduced, and the obtained antibody is high in correct conformation proportion and good in homogeneity and can be used for detecting human CD16 natural molecules through flow cytometry, IF, WB and the like.

The invention discloses a CD16 antibody and application thereof, the CD16 antibody comprises an antibody specifically binding to CD16A and an antibody strongly binding to CD16B but weakly binding to CD16A, the antibody specifically binding to CD16A can bind to CD16A but not binding to CD16B, and selectively activates NK cells and macrophages, so that adverse inflammatory reaction caused by cross reaction of CD16B is avoided; the antibody which is strongly combined with CD16B but weakly combined with CD16A has unique CD16B combining capacity, and can be used for enhancing the function of neutrophil and promoting immune complex removal, so that inflammatory response is regulated; the two antibodies can be developed into a plurality of drugs, have a wide application field, and provide a valuable technical platform for subsequent development of novel antibody drugs.

Provided herein is a recombinant polypeptide comprising: the transmembrane domain of FcεR1γ or CD3ζ, or a variant of the transmembrane domain that retains the ability to bind to NKp30, NKp46 and / or CD16, a co-stimulatory domain, and one or more immunoreceptor tyrosine-based activation motifs (ITAMs). Cells expressing the same and methods of treatment are also provided.

The invention relates to an anti-HER2 / CD16 / CCR4 trispecific antibody and a preparation method and application thereof.The trispecific antibody comprises a first antigen binding structural domain specifically bound with HER2, a second antigen binding structural domain specifically bound with CD16 and a third antigen binding structural domain specifically bound with CCR4, and the trispecific antibody can be bound with human HER2, CD16 and CCR4 at the same time, so that the trispecific antibody can be used for detecting human HER2, CD16 and CCR4. The anti-tumor activity is obviously enhanced. The trispecific antibody can more effectively activate NK cells, enhance ADCC effect, target HER2 positive tumor cells and remove immunosuppressive cells in a tumor microenvironment, and has the advantages of enhancing antitumor activity and overcoming immunosuppression.

The invention provides a DLL3 tri-specific antibody and application thereof, the tri-specific antibody comprises: a first binding region having CD3 binding activity, a second binding region having CD3 binding activity, and a third binding region having CD3 binding activity; a second binding region, the second binding region having a CD16 binding activity; and a third binding region, the third binding region having a DLL3 binding activity. The three-specific antibody can be combined with CD16 on NK cells, CD3 on T cells and DLL3 on the surfaces of tumor cells, and the NK cells and the tumor cells as well as the T cells and the tumor cells are promoted to be combined at the same time; the tri-specific antibody promotes formation of cell synapse between the two immune cells and the tumor cells, so that the activity of NK cells and T cells targeting the tumor cells is selectively redirected, various cancers can be effectively treated, and the application value is high.

The present application relates to a genetically modified immune cell for expressing a recombinant human CD16 protein. The recombinant human CD16 protein has additions, deletions and replacements of, or any combination of the additions, deletions and replacements of one or more amino acids as compared to a wild-type amino acid sequence, and has enhanced shear resistance.

The invention provides a B7H7 trispecific antibody and application thereof, the trispecific antibody comprises: a first binding region having CD3 binding activity; a second binding region, the second binding region having a CD16 binding activity; and a third binding region, the third binding region having a B7H7 binding activity. The trispecific antibody can be combined with CD16 on NK cells, CD3 on T cells and B7H7 on the surfaces of tumor cells, and the NK cells and the tumor cells as well as the T cells and the tumor cells are promoted to be combined at the same time; the tri-specific antibody promotes the formation of cell synapses between the two immune cells and tumor cells and thus selectively redirects the activity of NK cells and T cells to targeted tumor cells, and can effectively treat tumors.

The present invention provides a new NK cell culture medium, which belongs to the field of cell culture technology. It includes basal culture medium, serum, cytokines, antibiotics and auxiliary factors, wherein the basal culture medium is selected from RPMI-1640, DMEM or IMDM, the serum is fetal bovine serum or human serum, the cytokines include IL-2, IL-7, IL-15, IL-3, VEGF, GDNF, and the antibiotics are penicillin or streptomycin. The present invention introduces auxiliary factors containing paclitaxel, cocin, ethanolamine, hyaluronic acid, sodium selenite, L-ascorbic acid, pyrroloquinoline quinone, dihydroxypropylphylline, reduced glutathione, madecassoside, D-pantothenate calcium, and estradiol into the basal culture medium, which can significantly increase the proliferation multiples and cell viability of NK cells, and at the same time, promote the expression of CD56 and CD16 by progeny NK cells. Experimental results show that after a 16-day culture period, the present invention causes NK cells to proliferate 377.6 times, the cell viability reaches 98.2%, and the expression of CD16 + CD56 + The percentage of cell number reached 83.28%.

Described herein are pharmaceutical formulations comprising multi-specific binding proteins that bind to NKG2D, CD16, and epidermal growth factor receptor (EGFR); and therapeutic uses of the multi-specific binding proteins and pharmaceutical formulations thereof for treating a disease, for example, cancer, in a patient in need thereof.

The present application provides a bispecific antibody comprising a first antigen binding region that binds to human CD16 and a second antigen binding region that binds to human CD123, a single domain antibody that binds to human CD16, a pharmaceutical composition comprising the bispecific antibody or the single domain antibody, an antibody-natural killer cell (NK cell) conjugate, and uses thereof.

The invention discloses an antibody combination for typing CD123 + dendritic cells and application of the antibody combination. An antibody in the antibody combination consists of a CD45 antibody, a CD3 antibody, a CD20 antibody, a CD16 antibody, an HLA-DR antibody and a CD123 antibody. When the kit formed by the antibody combination provided by the invention is used for typing the CD123 + dendritic cells in the peripheral blood single cells, the quantity and the stability of the collected cells are remarkably improved.

The present invention relates to the field of virology. More specifically, the present invention provides compositions and methods useful for elimination of human immunodeficiency virus-1 (HIV-1) reservoir cells. In several embodiments, the single chain diabody binds human immunodeficiency virus 1 Envelope protein variable loops 1 and 2 (HIV-1 Env V1 / V2) and human type III Fcγ receptor (CD16). In a specific embodiment, the scDb comprises (a) an anti-HIV-1 Env V1 / V2 antigen binding fragment comprising a VH comprising or consisting of SEQ ID NO:1; (b) an anti-HIV Env V1 / V2 antigen binding fragment comprising a VL comprising or consisting of SEQ ID NO:5; (c) an anti-CD16 antigen binding fragment comprising a VH comprising or consisting of SEQ ID NO:17; and (d) an anti-CD16 antigen binding fragment comprising a VL comprising or consisting of SEQ ID NO:21.

The invention discloses a composition for efficiently amplifying high-purity NK cells and application of a kit, and relates to the technical field of cell culture. The antibody combination comprises a CD3 antibody, a CD16 antibody and an NKG2D antibody, wherein the mass ratio of the CD3 antibody to the CD16 antibody to the NKG2D antibody is (0.01-1): (2-8): (2-8). According to the composition for activating the NK cells, provided by the invention, the PBMC cells are cultured by using the composition, so that the purity and the number of the harvested NK cells can be greatly improved, the activity of the obtained NK cells is higher, and the motility rate can reach 90% or above. Therefore, the problems of low amplification purity and few amplification times of the NK cells are solved. In addition, the NK cell amplification method provided by the invention is simple and easy to implement, and is easy to popularize and apply.

The present invention relates to a pharmaceutical composition and cell therapy agent for the prevention or treatment of neurological diseases, comprising Schwann cell precursors (SCPs) or Schwann cells differentiated therefrom, derived from pluripotent stem cells (PSCs) or somatic cells, and natural killer (NK) cells as active ingredients. Specifically, the Schwann cell precursors (SCPs) express at least one selected from the group consisting of GAP43, SOX10, IGFBP2, and combinations thereof; the Schwann cells (SCs) express at least one selected from the group consisting of S100B, SOX10, and combinations thereof; and the natural killer (NK) cells express CD56 + CD16 + It is characterized by expressing at least one selected from the group consisting of the and combinations thereof.

This invention relates to the fields of biotechnology and targeted therapy, specifically to a recombinant construct, a recombinant oncolytic virus, its construction method, and its applications. The recombinant oncolytic virus CD19 BIKE is prepared by inserting exogenous genes for CD16 and CD19 into the genome of an oncolytic vaccinia virus. The antibodies secreted by this recombinant virus can not only bridge NK cells and CD19-positive B-cell lymphoma cells, increasing the contact between NK cells and tumor cells and their accumulation within tumor tissue, but also synergize with the tumor-lysing effect of the oncolytic virus, breaking down the stromal barrier, reshaping the tumor microenvironment, and enhancing therapeutic efficacy.

Provided herein are cells or populations of cells comprising a polynucleotide encoding a cleavage resistant CD16 polypeptide. Also provided herein are methods of suppressing the proliferation of tumor cells such as HER2+ and methods of treating cancers, such as HER2+ in a subject with populations of placental-derived natural killer cells or placental-derived T cells comprising a cleavage resistant CD16. The natural killer cells, such as CYNK cells, can be placental CD34+ cell-derived natural killer (NK) cells. The placental-derived T cells can be isolated from cord blood or from placental perfusate.

The invention discloses an activity-enhanced NK immune cell and an application thereof in cancer treatment. The NK cell stably expresses an anti-PD-L1 / CD16 coupled bispecific antibody through a lentiviral vector, and the antibody is formed by connecting an anti-PD-L1 single-domain antibody and an anti-CD16 single-domain antibody in series through a flexible connecting peptide (G4S). The NK immune cell with enhanced activity has three functions at the same time: tumor cells are targeted through anti-PD-L1 sdAb, and a PD-1 / PD-L1 immunosuppression pathway is blocked; the ADCC effect of the NK cell itself is activated through the anti-CD16 sdAb; by virtue of the small molecular weight characteristic of the single-domain antibody, the penetrating ability to solid tumors is remarkably enhanced. An in-vitro experiment shows that the killing rate of the sdAb-NK cell on various PD-L1 positive tumor cells is greatly improved compared with that of an unmodified NK cell, and the tumor penetrating power is improved by 3.7 times. In-vivo animal experiments show that the tumor inhibition rate is up to 84.8%, no obvious toxicity is shown, and a new efficient and safe strategy is provided for immunotherapy of solid tumors.