41 results about "Premalignant Cell" patented technology

CD20 is a transmembrane protein of the tetra-spanin family expressed on the surface of B-cells and has been found on B-cells from peripheral blood as well as lymphoid tissues. CD20 expression persists from the early pre-B cell stage until the plasma cell differentiation stage. Conversely, it is not found on hematopoietic stem cells, pro-B cells, differentiated plasma cells or non-lymphoid tissues. In addition to expression in normal B-cells, CD20 is expressed in B-cell derived malignancies such as non-Hodgkin's lymphoma (NHL) and B-cell chronic lymphocytic leukemia (CLL). CD20 expressing cells are known to play a role in other diseases and disorders, including inflammation. The present invention includes anti-CD20 antibodies, forms and fragments, having superior physical and functional properties; immunoconjugates, compositions, diagnostic reagents, methods for inhibiting growth, therapeutic methods, improved antibodies and cell lines; and polynucleotides, vectors and genetic constructs encoding same.

The invention discloses an NK cell in-vitro amplification composition and an NK cell amplification method and relates to an NK cell amplification composition and an amplification method. The invention aims at solving the problems of low multiple, low purity, long amplification period and high cost of current NK cell amplification. The composition comprises NK amplification trophocyte, IL-2, inactivated autologous plasma and GT-T551 H3 culture medium. The amplification method comprises the following steps: re-suspending PBMCs with the GT-T551 H3 culture medium; transferring into a culture bottle; adding the IL-2, NK amplification trophocyte and autologous plasma; and performing in-vitro co-culture for 15 days, wherein the NK amplification trophocyte is supplemented in the 7th day of the culture, the culture medium is supplemented in the culture process, and the cells are collected for detection in the 13th and 15th days of the culture. The composition can remarkably promote mass amplification of NK cells to meet the needs of clinical application; and the obtained NK cells have relatively high purity, the cost is saved, and the killing activity is improved. The NK cell in-vitro amplification composition and NK cell amplification method are applied to the amplification of NK cells.

A novel family of human mitochondrial RNAs, referrred to as chimeric RNAs, which are differentially expressed in normal, pre-cancer and cancer cells, are described. Oligonucleotides targeted to the chimeric RNAs are provided. The described oligonucleotides or their analogs can be used for cancer diagnostics and cancer therapy as well as for research. In one embodiment of this invention, these oligonucleotides hybridize with the sense or with the antisense mitochondrial chimeric RNAs, and the result of the hybridization is useful to differentiate between normal proliferating cells, pre-cancer cells and cancer cells. In another embodiment of the invention, the compositions comprise oligonucleotides that hybridize with the human chimeric RNAs resulting in cancer cell and pre-cancer cell death, while there is no effect in normal cells, constituting therrefore, a novel approach for cancer therapy.

A novel family of human mitochondrial RNAs, referred to as chimeric RNAs, which are differentially expressed in normal, pre-cancer and cancer cells, are described. Oligonucleotides targeted to the chimeric RNAs are provided. The described oligonucleotides or their analogs can be used for cancer diagnostics and cancer therapy as well as for research. In one embodiment of this invention, these oligonucleotides hybridize with the sense or with the antisense mitochondrial chimeric RNAs, and the result of the hybridization is useful to differentiate between normal proliferating cells, pre-cancer cells and cancer cells. In another embodiment of the invention, the compositions comprise oligonucleotides that hybridize with the human chimeric RNAs resulting in cancer cell and pre-cancer cell death, while there is no effect in normal cells, constituting therefore, a novel approach for cancer therapy.

The invention discloses a method for preparing a heat-resisting attenuated virus live vaccine for goatpox by using a BHK21-C13 passage cell. Based on the existing attenuated virus AV41 for goatpox, which has excellent immunogenicity, the method comprises the following steps: culturing 25-28th generations of virus solution with a heterologous BHK21 cell, and then adding an appropriate heat-resisting freeze-drying protective additive to obtain the heat-resisting attenuated virus live vaccine for goatpox. The passage cell vaccine used by the method is superior to the primary cell vaccine in the aspects of virus yield, homogeneity, purity and the like, not only guarantees the effective level of the vaccine and has a better heat-resisting protection effect, but also can ensure the safety of a purebred pregnant goat. As the existing BHK21 cell adopts a mature suspension culture process, the exploration of various parameters of adherent culture lays a solid foundation for the next promotion of suspension process.

The invention relates to an in-vitro culture method for increasing human Vdelta2 T cell amplification efficiency and an application thereof. The method is characterized in that peripheral blood mononuclear cells are separated, a medium (AIMU+10%FBS)is used to prepare a cell suspension, zoledronate is added for in-vitro stimulation; after one-day stimulation of zoledronate, cytokine rhIL-2 is added every day; the human peripheral blood mononuclear cells are cultured for 72-96 hours, half amount or 2 / 3 mount of nutrient solution can be replaced when the nutrient solution turns yellow; half amount of nutrient solution can be replaced when the nutrient solution turns yellow after 96 hours; when culture is carried out in 11th-14th day, when the amplified Vdelta2 T cell frequency and absolute amount reach the requirements, the cells can be acquired. The Vdelta2 T cell purity can reach more than 90%. The method has the advantages of simple culture method, the required peripheral blood amount is little, induction cell quantity is more, and the amplification efficiency is high, Vdelta2 T cell is expected to increase curative effect of for liver cancer immunity treatment, and has good application potential.

The present invention provides a culture method for increasing amplification efficiency and activity of Vgamma9Vdelta2T cells, wherein IL-2 and zoledronic acid are combined with a tyrosine kinase inhibitor dasatinib to carry out in vitro Vgamma9Vdelta2T cell induction production to obtain the Vgamma9Vdelta2T cells with high induction efficiency and high activity. According to the present invention, the method has characteristics of simple and easy performing induction culture process, short period, low cost, high induction efficiency and good repeatability, the cells obtained through induction has enhanced activity, the tumor immunotherapy effect of the current Vgamma9Vdelta2T cells is expected to be increased with the number and the function of the induction-cultured cells, and good application potential is provided.

Particular members of the multisubunit immune recognition receptor (MIRR) family of receptors, specifically, the B cell antigen receptor (BCR), the pre-B cell receptor (pre-BCR), the pro-B cell receptor (pro-BCR), Ig Fc receptors (FcR), and NK receptors, can be physically uncoupled from their associated transducers. The invention describes regulatory compounds and methods for mimicking such dissociation / destabilization for the purposes of receptor desensitization and for treatment of conditions in which receptor desensitization or alternatively, enhanced or prolonged receptor sensitization, is desirable. Compounds and methods for enhancing or prolonging receptor sensitization are also disclosed, as are methods for identifying regulatory compounds suitable for use in the present methods.

Compositions and methods for minimizing antibody disulfide bond reduction are described. In one aspect, a composition is provided for culturing mammalian host cells to express an antibody including an anti-reduction agent that minimizes reduction of a disulfide bond in the antibody or fragment thereof. In some other aspects, methods for minimizing disulfide bond reduction; increasing production of an antibody or fragment thereof with intact native disulfide bonds; increasing a ratio of non-reduced to reduced antibody or fragment thereof; producing a therapeutic antibody or fragment thereof by adding a sufficient amount of an anti-reduction agent to a cell culture media, pre-harvest cell culture fluid, or harvest cell culture fluid are described. In another aspect, minimizing disulfide bond reduction in an antibody or fragment thereof culturing the host cell in a concentration of at least about 20% O2 is described.

CD20 is a transmembrane protein of the tetra-spanin family expressed on the surface of B-cells from peripheral blood as well as lymphoid tissues. CD20 expression persists from the early pre-B cell stage until the plasma cell differentiation stage. In addition to expression in normal B-cells, CD20 is expressed in B-cell derived malignancies such as non-Hodgkin's lymphoma (NHL) and B-cell chronic lymphocytic leukemia (CLL). The present invention includes anti-CD20 antibodies and antigen-binding fragments thereof comprising a light chain variable region and a heavy chain variable region, wherein the CDR-L1, CDR-L2, and CDR-L3 of said light chain variable region comprise the amino acid sequences of SEQ ID NOs: 17-19, respectively, and wherein the CDR-H1, CDR-H2, and CDR-H3 of said heavy chain variable region comprise the amino acid sequences of SEQ ID NOs: 20-22, respectively.

The invention relates to a method for evaluating CAR-T killing activity in vitro. IN the prior art, the existing CAR-T cell in-vitro killing activity detection method relates to isotope safety, operation is tedious and other problems exist. Based on the problems in the prior art, in order to conveniently and simply realize the detection of in vitro killing activity of CAR-T cells, a cell strain capable of evaluating the CAR-T killing function in vitro is constructed, and a method for detecting the CAR-T in-vitro killing activity through RTCA is provided, wherein the CAR-T cell killing activitycan be efficiently, sensitively and continuously detected in vitro through the method so as to provide the guarantee for the subsequent treatment effect of CAR-T cells.

The invention provides a biotin-avidin-lentiviral expression vector and a CAR-T (chemric antigen receptor T) cell preparation method, and aims at solving the defects of single targeting ability, complex preparation, time-consuming and labor-consuming properties and poor economical efficiency of traditional CAR-T. The method comprises the following steps of constructing an avidin-CAR lentivirus package carrier; performing virus packaging; and performing T cell separation, activation, co-stimulation structural domain gene transduction and amplification, thereby preparing CAR-T cells. On the basis of a general CAR-T treatment system, a molecular imaging detection means is introduced, a cell-based visualization treatment scheme is provided, and a CAR-T cell treatment process can be dynamically monitored in vivo in real time, so that a situation that the current CAR-T cell treatment process is lack of effective monitoring is solved. The biotin-avidin-lentiviral expression vector has the advantages of simplicity in preparation, diversity in selection, extensive targets and visualization on treatment process, and suitability for treatment of tumor patients.

The invention discloses an ROBO1 CAR-NK cell carrying a suicide gene as well as a preparation method and application of the ROBO1 CAR-NK cell. In order to improve the safety and controllability of theCAR-NK therapy, a suicide gene switch element is integrated into a genome through a lentiviral transfection technology on the basis of the current ROBO1 CAR-NK cell to form the CAR-NK with the suicide gene. By adding the suicide gene, the CAR-NK cells can be better controlled, and the clinical safety is further improved.

The invention discloses an epinephelus lanceolatus head kidney cell line as well as a construction method and application thereof. The ELHK cell line is preserved in the Guangdong Microbial Culture Collection Center (GDMCC) on December 04, 2020, the address is the 5th floor of No.59 building, No.100, Middle Xianlie Road, Guangzhou City, Guangdong Province, the postal code is 510070, and the preservation number is GDMCC No: 61340. The Epinephelus lanceolatus head kidney cell line-ELHK cell line is obtained, the growth state is good, cell proliferation is stable, the cell morphology takes a fibroblast sample as the main morphology, continuous passage can be achieved (at present, the cells have already been transferred to more than 120 generations), ultralow-temperature cryopreservation and recovery can be achieved. The establishment of the cell line lays a foundation for related research of grouper germplasm resource preservation.

Disclosed is a new class of conjugated virus-like particles (VLPs). These conjugated VLPs bind a wide variety of tumors and comprise epitopes recognized by a prior T cell immune response already existing in a host. These epitopes are derived from pathogens or previous vaccinations (such as early childhood vaccines). This provokes the body's pre-existing cytotoxic immunity obtained through previous infection or previous childhood vaccination to be redirected to the tumor cells for the elimination of cancer, and form long-term anti-tumor immunity. The described conjugated VLPs are useful for tailoring a broad range of tumors towards a response from existing immunity circumventing the need to identify tumor antigens or generate tumor-specific immune responses. Importantly, the compositions and methods described herein broadens opportunities for treatment for all cancer types in subjects who previously had un-targetable cancers due to various technological and biological limitations of currently available immuno-therapeutic drugs.

The invention provides a gelatin cell scaffold with a controllable porous structure and a preparation method thereof. The preparation method comprises the following steps: firstly, preparing uniform latex emulsion through an emulsion process; secondly, adding a cross-linking agent into the latex emulsion to prepare gelatin microspheres; thirdly, bonding the gelatin microspheres in a crosslinking manner in a demulsification process to form gel; lastly, performing freeze drying on the gel to obtain the gelatin cell scaffold. The gelatin cell scaffold with the controllable porous structure is prepared by bonding the gelatin microspheres of different particle sizes, so that the problem that the conventional cell scaffold is non-uniform in particle size and is uncontrollable in size is solved effectively. The gelatin cell scaffold provided by the invention has high biocompatibility, biodegradability and higher mechanical performance. Moreover, the preparation method is green, is environmentally friendly, is easy to operate, and is feasible. Thus, the gelatin cell scaffold provided by the invention has a good market application prospect in the field of tissue engineering.

The present invention provides a cell-penetrating peptide-pre-B cell leukemia transcription factor 1 fusion protein and preparation method and application thereof, and relates to the technical field of biomedicine; PBX1 transcription factor is efficiently introduced into stem cells by means of a CPP and PBX1 fusion expression mode by making use of the cell-penetrating characteristic that CPP can carry exogenous proteins to enter cells, so that self-renewing capacity of the stem cells is retained, the proliferation of the stem cells is promoted, the senescence of the stem cells is delayed, anda new solution is provided for the in-vitro large-scale preparation of the stem cells.

The invention discloses an infection enhancing culture medium and method for improving the cell lentivirus infection rate, and relates to the field of MDA-MB-231 cell transfection. The enhanced infection culture medium is used for improving the lentivirus infection rate of the MDA-MB-231 cell, and the enhanced infection culture medium is prepared from the following components: a Leibovitz's L-15 culture medium, an RPMI-1640 culture medium, fetal calf serum, non-essential amino acid and an epidermal growth factor; according to the volume percentage of the total raw materials, the addition amount of the fetal calf serum is 10%, and the addition amount of the non-essential amino acid is 1%; the addition amount of the epidermal growth factor is 2-3ng / ml. By adopting the enhanced infection culture medium disclosed by the invention, the MDA-MB-231 cells can be normally cultured in the conventional environment of 37 DEG C and 5% CO2, and the infection efficiency of the lentivirus on the MDA-MB-231 cells is more remarkably improved, so that the gene editing efficiency of the MDA-MB-231 cells becomes higher, and the problem that the lentivirus infection rate of the MDA-MB-231 cells is low at present is solved.

The invention belongs to the technical field of medicines, and discloses a phosphate ester derivative of herba epimedii as well as a preparation method and application of the phosphate ester derivative. The chemical structural formula of the phosphate ester derivative of the herba epimedii is represented by the formula (1), wherein R1 is selected from groups defined in the specification, or R2 is selected from H or R1, and R3 is selected from H or a group defined in the specification. According to the phosphate ester derivative of the herba epimedii, the cytotoxicity of the partially modified derivative is remarkably enhanced compared with that of an unmodified derivative, and meanwhile the phosphate ester derivative of the herba epimedii has the anti-osteoporosis effect. The phosphate ester derivative of the herba epimedii can be applied to the field of preparation of drugs for prostate cancer cells, anti-osteoporosis health-care products, pharmaceutical excipients or drugs.

Compositions and methods for minimizing antibody disulfide bond reduction are described. In one aspect, a composition is provided for culturing mammalian host cells to express an antibody including an anti-reduction agent that minimizes reduction of a disulfide bond in the antibody or fragment thereof. In some other aspects, methods for minimizing disulfide bond reduction; increasing production of an antibody or fragment thereof with intact native disulfide bonds; increasing a ratio of non-reduced to reduced antibody or fragment thereof; producing a therapeutic antibody or fragment thereof by adding a sufficient amount of an anti-reduction agent to a cell culture media, pre-harvest cell culture fluid, or harvest cell culture fluid are described. In another aspect, minimizing disulfide bond reduction in an antibody or fragment thereof culturing the host cell in a concentration of at least about 20% O2 is described.

Disclosed is a new class of conjugated virus-like particles (VLPs), These conjugated VLPs bind a wide variety of tumors and comprise epitopes recognized by a prior T cell immune response already existing in a host. These epitopes are derived from pathogens or previous vaccinations (such as early childhood vaccines). This provokes the body's pre-existing cytotoxic immunity obtained through previous infection or previous childhood vaccination to be redirected to the tumor cells for tire elimination of cancer, and form long-term anti-tumor immunity. The described conjugated VLPs are useful for tailoring a broad range of tumors towards a response from existing immunity circumventing the need to identify tumor antigens or generate tumor-specific immune responses, importantly, the compositions and methods described herein broadens opportunities for treatment for ail cancer types in subjects who previously had un-targetable cancers due to various technological and biological limitations of currently available immuno-therapeutic drugs.

The invention relates to a DNA transmethylase Dnmt3b gene deficient type CHO (Chinese hamster ovary) cell line, a preparation method and application thereof and a recombinant protein expression system, and belongs to the technical field of genetic engineering. CHO cell Dnmt3b gene is knocked off by CRISPR / Cas9 gene editing technique so as to obtain the Dnmt3b gene deficient type CHO cell line is attained; the Dnmt3b gene deficient type CHO cell line can evidently increase the expression level and expression stability of a target gene in CHO cells, and the problem can be solved that existing CHO cell expression systems have low expression level and expression instability; upon expression of recombinant Adalimumab with the cell line herein, it is discovered that the expression level of the recombinant Adalimumab is increased evidently. Therefore, the cell line of the invention is widely applicable to the expression of target proteins.

Particular members of the multisubunit immune recognition receptor (MIRR) family of receptors, specifically, the B cell antigen receptor (BCR), the pre-B cell receptor (pre-BCR), the pro-B cell receptor (pro-BCR), Ig Fc receptors (FcR), and NK receptors, can be physically uncoupled from their associated transducers. The invention describes regulatory compounds and methods for mimicking such dissociation / destabilization for the purposes of receptor desensitization and for treatment of conditions in which receptor desensitization or alternatively, enhanced or prolonged receptor sensitization, is desirable. Compounds and methods for enhancing or prolonging receptor sensitization are also disclosed, as are methods for identifying regulatory compounds suitable for use in the present methods.

The invention provides a simple and convenient method for preparing universal immune cells and an application of the universal immune cells. According to the method, allogeneic immune cells, specificcell mitogens, cell factors and immunologic adjuvants are placed in a liquid cell culture medium to be co-cultured in a culture container, so that universal immune cell culture with high immunocompetence is obtained. The method is simple, convenient and easy to operate, the allogeneic immune cells are adopted, the quality of immune cell preparations can be improved, the cost of cell therapy is reduced, and the safety of the cell therapy is improved. In addition, a large number of clinical long-term practical applications prove that the universal immune cells are safe, reliable, durable and effective, and have no rejection reaction. Therefore, the universal immune cells can be used as maternal cells of other types of universal immune cell preparations, such as CAR-T and TCR-T, and can be used in the fields of adoptive cellular immunotherapy and the like. According to the invention, the problems of high cost, poor safety, low efficiency and the like of existing cellular immunotherapy aresolved, and a new way is provided for application and popularization of the cellular immunotherapy.

Nanostraws and to methods of utilizing them in order to deliver biologically relevant molecules such as DNA, RNA, proteins etc., into non-adherent cells such as immune cells, embryos, plant cells, bacteria, yeast etc. The methods described herein are repeatedly capable of delivering biologically relevant cargo into non-adherent cells, with high cell viability, dosage control, unaffected proliferation or cellular development, and with high efficiency. Among other uses, these new delivery methods will allow to scale pre-clinical cell reprogramming techniques to clinical applications.

The invention relates to the technical field of genetic engineering, in particular to a CHO cell line, a construction method, a recombination protein expression system and an application. The APRT gene defective Type CHO cell line is used for constructing the recombination protein expression system, and no matter whether G418 screening pressure exists or not, the expression and maintaining rate ofa target protein eGFP in the recombination protein expression system constructed with 60th generation APRT gene defective type CHO cells is far higher than that of eGFP in normal recombination CHO cells. When the G418 screening pressure does not exist, the expression level of recombination vitronectin in the 30th generation cells of the recombination protein expression system constructed by the CHO cells is notably higher than that of corresponding target proteins in normal CHO cells. The recombination protein expression system can notably increase the expression level and long-term expression stability of the target gene in the CHO cells, and the problem that present CHO cell expression system is unstable in expression, can be solved.

The invention discloses application of a hemopoietic pill in the preparation of a medicine for preventing or repairing medullary cell damage caused by environmental pollutions, wherein the environmental pollutions are lead pollution, cigarette pollution, benzene pollution, dimethylbenzene pollution and formaldehyde pollution, and the preventing or the repairing of the medullary cell damage refers to the restraint of the proliferation of a precancerous cell. In the invention, the functions that the hemopoietic pill resists to the teratogenesis caused by the environmental pollutions are inspected on aspects of the change of the micronucleus rate of blood lymphocytes and marrow, the elimination capability of free radicals, the restraint of the proliferation of the precancerous cell, and the like, and the wider foreground is opened up for the clinical application of the hemopoietic pill.

The invention relates to the technical field of medicine, in particular to a cell or stem cell cryopreservation preparation method which comprises the following steps: step 1, protecting stem cells through a protective agent; step 2, pre-freezing the stem cells; step 3, carrying out primary drying on the pre-frozen stem cells; and 4, carrying out secondary drying on the stem cells subjected to primary drying. The freeze-dried cells can be preserved at normal temperature through the protective agent, the performance is stable, the preservation time is long, transportation is convenient, the preservation cost is low, and the limitation of current cell preservation is solved.

Methods for inhibiting the motility or proliferation of premalignant and malignant cells are provided. Methods for treating a malignancy of the head and neck and for treating a malignancy of the lung are also provided.