184 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.

Combination therapy of a cancer with a multi-specific binding protein that bind a tumor associated antigen, the NKG2D receptor, and CD16, in combination with a second anti-cancer agent are described. Also described are pharmaceutical compositions of the multi-specific binding protein, and therapeutic methods useful for the treatment of cancer in combination with a second anti-cancer agent.

The invention belongs to the technical field of culture media, and particularly relates to a culture medium for promoting in-vitro amplification of NK cells, the culture medium for in-vitro amplification of the NK cells comprises a basic culture solution, a CD16 antibody, IL2, IL12 and IL21, the basic culture solution is DMEM, and the culture medium for in-vitro amplification of the NK cells is obtained by adding autologous plasma, the CD16 antibody, the IL2, the IL12 and the IL21 on the basis of the basic culture solution. Wherein the addition amount of the autologous plasma is 10%-15%, and the final concentrations of the CD16 antibody, IL2, IL12 and IL21 are 1 [mu] g / mL to 20 [mu] g / mL, 1000 IU / mL to 2000 IU / mL, 20 ng / mL to 40 ng / mL and 5 ng / mL to 15 ng / mL. According to the method, the culture medium is optimized, the single NK cell is resuspended by using the complete culture medium, the single NK cell is added into the CD16 antibody coated cell culture plate and cultured for 21 days or more, the method promotes in-vitro amplification of the NK cell, and the obtained NK cell has the advantages of being high in purity and large in number, saving the cost and improving the killing activity.

The invention belongs to the technical field of immune cell culture, and particularly relates to an in-vitro amplification culture method of NK cells. The amplification method comprises the steps of CD16 pretreatment, plasma inactivation, lymphocyte separation and multi-stage fluid infusion. The preparation method comprises the following steps: pretreating CD16 in a T75 culture flask, promoting attachment and activation of NK cells, simulating a natural activation state, improving proliferation efficiency, treating whole blood and inactivating plasma to eliminate immunosuppressive factors, reserving nutritional ingredients, supporting amplification of the NK cells, enriching the NK cells through a lymphocyte separating medium, removing impurities and improving purity, mixing an RPMI-1640 culture medium with a solution A, and culturing for 12-24 hours at a high temperature; oK432 and IL-15 are added, the microenvironment is optimized, growth of the NK cells is remarkably promoted, liquid supplementing operation and bottling and bagging are carried out for multiple times, sufficient space and nutrition are provided, the cells are gradually transferred into a serum-free culture medium, and the cell safety and clinical application potential are improved.

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.

The present application relates to genetically modified immune cells to express a recombinant human CD16 protein having an enhanced anti-shear ability with the presence of addition, deletion, substitution, or any combination of addition, deletion, substitution of one or more amino acids compared to a wild-type amino acid sequence.

The invention relates to an antibody composition, a kit and a system for rapidly screening acute promyelocytic leukemia. The antibody composition comprises a first group of antibodies, a second group of antibodies and a third group of antibodies, the first group of antibodies comprises a CD71 antibody, a CD33 antibody, a CD117 antibody, a CD34 antibody, an HLA-DR antibody, a CD19 antibody, a CD10 antibody, a CD7 antibody and a CD45 antibody; the second group of antibodies comprises a CD38 antibody, a CD56 antibody, a CD16 antibody, a CD34 antibody, a CD13 antibody, a CD11b antibody, a CD64 antibody, a CD15 antibody and a CD45 antibody; the third group of antibodies comprises a CD9 antibody, an MPO antibody, a CD117 antibody and a CD45 antibody; and each antibody is a monoclonal antibody marked with a detection marker. The antibody composition comprises a CD117 + CD33 + region as a main target cell population, and covers three groups of antibodies for identifying acute promyelocytic leukemia, such as CD33, CD34, CD64, CD13, CD9 and MPO, the acute promyelocytic leukemia can be rapidly, simply and conveniently detected by adopting the antibody composition, the detection sensitivity is high, the specificity is high, the accuracy rate is high, and the antibody composition can be widely applied to detection of acute promyelocytic leukemia. The acute promyelocytic leukemia patient can be accurately and quickly identified, and early diagnosis and early treatment are realized.

Multi-specific binding proteins that bind to and kill human cancer cells are described, as well as pharmaceutical compositions and therapeutic methods useful for the treatment of cancer. The cancer can be B-cell maturation antigen (BCMA)-expressing cancer. The multi-specific binding proteins provided herein exhibit high potency and maximum lysis of target cells compared to anti-BCMA monoclonal antibodies.

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.

Multi-specific binding proteins that bind NKG2D receptor, CD 16, and a tumor-associated antigen selected from c-MET, KIT, F3, IGF1R, Lewis Y, MUC13, MUC4, MCAM, LRRC32, sialyl-Tn, gpA33, GD3, GM2, EPHA3, TNFRSF10A, TNFSF11, CD74, and PMEL are described, as well as pharmaceutical compositions and therapeutic methods useful for the treatment of cancer.

Multi-specific binding proteins that bind to and kill human cancer cells expressing epidermal growth factor receptor 2 (HER2 or ErbB2), but does not kill non-cancerous healthy human cells expressing HER2 are described, as well as pharmaceutical compositions and therapeutic methods useful for the treatment of HER2 expressing cancer. The invention also relates to multi-specific binding proteins that trigger CD8+ T cell killing of tumor cells.

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.

Multi-specific binding proteins that bind to and kill human cancer cells expressing CD33 (Siglec-3) are described, as well as pharmaceutical compositions and therapeutic methods useful for the treatment of CD33 expressing cancer. The invention relates to multi-specific binding proteins that bind to human cancer cells expressing CD33 and exhibit high potency and maximum lysis of target cells compared to anti-CD33 monoclonal antibodies. The multi-specific binding proteins comprise a CD33-binding domain, an NKG2D-binding domain and a CD16-binding domain.

Provided are genetically modified NK cells expressing a chimeric antigen receptor targeting an EGFR superfamily receptor. The CAR can comprise an intracellular domain of FcεRIγ and further recombinant proteins expressed by the genetically modified NK cells are CD16, autocrine growth stimulating cytokines, and optionally one of IL-12, a TGF-beta trap, or a homing receptor. Also described are methods for treating a patient having or suspected of having a disease that is treatable with NK-92 cells, such as cancer, comprising administering to the patient the genetically modified NK cells.

This disclosure relates to methods of treating cancer using multi-specific binding proteins that bind NKG2D, CD16 and a tumor-associated antigen such as HER2. Provided are uses of the multi-specific binding protein in combination with a corticosteroid to reduce the risk of infusion-related reactions. Also provided are uses of the multi-specific binding protein in treating cancer that has low or moderate HER2 expression level. The present disclosure also relates to pharmaceutical formulations comprising the multi-specific binding proteins.

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.

Provided herein are compositions of NK-92® cells that express a CD19 CAR, CD16 and IL2, and the method of using these cells to and treat cancer in a patient.

An NK cell showing higher cytotoxic activity is provided. An object of the present invention is to provide a pharmaceutical composition for NK cell therapies expected to be highly effective. The present invention provides an NK cell having the following characteristics of (1) and (2) or a population thereof: (1) the NK cell is CD16-positive, highly expresses CD56, and is CD57-negative, and (2) the NK cell is NKG2C-positive, is NKG2A-negative or lowly expresses NKG2A, and is CD94-positive. The present invention also provides a pharmaceutical composition containing a population of such NK cells, and a therapeutically effective amount of antibodies.

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.

To provide methods for preparing novel immunoregulatory macrophage cells which are useful in the treatment of different immunological and non-immunological diseases and conditions; resulting macrophage cells; and pharmaceutical compositions comprising macrophage cells.SOLUTION: A method for preparing an immunoregulatory macrophage cell is provided, the method comprising: (a) isolating CD14 positive monocytes from a blood sample of a subject; (b) culturing the monocytes in a gas-permeable bag in a culture medium containing (i) M-CSF and / or GM-CSF, and (ii) a CD16 ligand; (c) contacting the cells with IFN-γ; and (d) obtaining the immunoregulatory macrophage cell from the culture medium.SELECTED DRAWING: Figure 10

The name of the invention is NK zygote molecules and methods of use thereof. Compositions for activating NK cells to stimulate an immune response for the treatment of cancer and other disorders are provided. In one embodiment, the present invention provides a compound comprising an NK binding domain that binds to CD16; an NK activation domain operably linked to the NK binding domain; and a targeting domain selectively binding to a target cell and operably linked to the NK activating domain and the NK binding domain, wherein the targeting domain binds to CLEC12A.

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.

The invention belongs to the field of biomedicine, and particularly relates to a detection method, an evaluation method and application of umbilical cord blood-derived natural killer cells. The detection method disclosed by the invention is used for carrying out related detection and data analysis on the natural killer cells from umbilical cord blood, is high in pertinence and can be used as a detection method for delivering the NK cells from a warehouse. According to the evaluation method, the relationship between the NK function related protein and the cell killing ability is determined, the killing ability of the NK cells is judged by detecting functional protein indexes CD3, NKG2A, CD56, CD16, NKP46 and CD107A of the natural killer cells and calculating NK scores of samples based on detection results, the sensitivity and specificity are high, and the method can be used for evaluating the killing ability of the NK cells.

The invention discloses an NK cell culture solution and a culture method thereof. The culture solution comprises an amplification culture solution and an induction activation culture solution, wherein the amplification culture solution is based on a serum-free culture medium and is added with cell factors such as IL-2, IL-12, IL-15 and IL-18, and the induction activation culture solution is based on the amplification culture solution and is added with induction activation factors such as OK432, CD16-alpha, a pseudomonas aeruginosa injection, alpha-galactosylceramide and CD137L. The induction activation factor CD16-alpha added in the NK cell culture fluid can activate NK cells independently or in combination with OK432, a pseudomonas aeruginosa injection, alpha-galactosylceramide and the like, so that the NK cells grow quickly, and the CD3-C56 + ratio of the NK cells can be increased; after the CD16-alpha is combined with the CD137L, the activated NK cells can be further activated, so that the activity of the NK cells is higher, and the killing effect is stronger.

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

The invention belongs to the technical field of biological medicine, and particularly relates to a frozen umbilical cord blood NK cell amplification method which comprises the following steps: S1, umbilical cord blood NK cell separation; s2, performing serum-free culture; s3, performing dynamic amplification; and S4, performing cryopreservation and resuscitation verification. According to the invention, after unfreezing, a culture medium containing a TGF-beta inhibitor (Galunissertib) is firstly used for pretreatment for 24 hours, so that an inhibitory signal is blocked, and the survival rate is increased. The dynamic amplification adopts three-stage metabolism regulation: stage 1: in a low-oxygen environment, using a culture medium containing SCF + FLT3L + IL-7 to promote activation of NK cells in a stationary period; stage 2: under a normal oxygen condition, switching to IL-12 + IL-18 + IL-21, and adding an antioxidant NAC to reduce ROS accumulation; and stage 3: pulse-type stimulation: adding CD16 cross-linked magnetic beads every 48 hours to enhance cytotoxicity.

A recombinant NK cell, in particular a recombinant NK-92 cell, expresses a chimeric antigen receptor (CAR) having an intracellular domain of Fc [epsilon] RI [gamma]. Notably, the CAR constructs having the intracellular domain of Fc [epsilon] RI [gamma] have a significantly prolonged expression duration and a significantly prolonged cytotoxicity over time. The CARs can be expressed from RNA and DNA, preferably as tricistronic constructs that further encode CD16 and cytokines to confer autocrine growth support. Advantageously, such constructs also enable high levels of transfection and expression of recombinant proteins and provide convenient selection markers to promote rapid production of recombinant NK / NK-92 cells.

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.