40 results about "Monoclonal antibody production" patented technology

Production of recombinant human rabies virus mono-cloning antibody NM57 is stable and has high purity and excellent specificity. It can be used to prevent rabies and substitute ERIG and HRIG.

The present invention provides for an improved process to obtain substantial amount of monoclonal antibodies with desired profile of charged variants. The process involves initially culturing e mammalian cells at a suitable temperature and subsequently reducing the temperature and optionally by simultaneous addition of suitable amino acid(s) during production of the desired molecule. The present invention provides also provides for an antibody having desired profile of glycans prepared with said with improved process.

A method of increasing animal cell growth and monoclonal antibody production in an animal cell or cell culture includes the use of ultrasound at a frequency greater than about 1 MHz.

The invention discloses a preparation method of a novel coronavirus nucleocapsid protein monoclonal antibody. The preparation method comprises the following steps: S1, synthesizing polypeptides; S2, preparing an immunogen; S3, preparing a screening antigen; S4, immunizing animals; S5, fusing cells; S6, screening and detecting; S7, establishing a stable cell strain; S8, producing the monoclonal antibody; S9, coupling the antibody; and S10, screening paired antibodies. The polypeptides are synthesized by selecting different antigen epitopes at two ends of N protein respectively and are used as antigens; the prepared monoclonal antibody can realize direct pairing; in a polypeptide synthesis process, a connecting bridge is added and steric hindrance is reduced, so that the antigen epitopes aresufficiently exposed on the surface of carrier protein and the antigens can be rapidly identified; recombinant protein is selected as a detection antigen, so that the risk that the screened antibodyonly identifies polypeptide antigens and does not identify the N protein is avoided; and whether a sample contains novel coronaviruses or not can be directly detected by detecting the content of the Nprotein.

This invention provides improved methods for production of monoclonal antibodies against a protein of interest. The present methods are based on immunization of an animal with a fusion protein between a protein of interest and a Th2 cytokine such as IL-4, IL-5, IL-13 and IL-31.

The invention discloses a method for producing a monoclonal antibody by using hybridoma. The method comprises the following steps: placing resuscitated hybridoma in a first cell culture medium for performing sub-culture and enlarge culture, wherein the volume fraction of serum in the first cell culture medium is larger than 10 percent; transferring the hybridoma into a second cell culture medium for performing enlarge culture, wherein the volume fraction of serum in the second cell culture medium is less than or equal to 10 percent; transferring the hybridoma into a serum-free cell culture medium for culturing, and performing monoclonal antibody production. By adopting the method, the first cell culture medium is used for performing eutrophic culture, and the second cell culture medium isused for performing uniform group culture, so that the concentration and volume of hybridoma of different batches can keep consistent at the start of antibody production, the whole production flow canbe widely applied to different hybridoma plants, and the growth period of the antibody is stable. The monoclonal antibody produced in the serum-free cell culture medium has high product quality.

The present invention discloses an anti-human-liver-cancer-stem-cell monoclonal antibody, and particularly relates to a new monoclonal antibody capable of inhibiting human liver cancer stem cells in a targeting manner, a hybridoma cell line secreting the monoclonal antibody, and applications of the monoclonal antibody in preparation of drugs for treatment of human liver cancer recurrence and drug resistance. According to the present invention, the monoclonal antibody can significantly inhibit the self-renewal, the invasion and the drug resistance of human liver cancer stem cells in vitro, can significantly inhibit the growth of human liver cancer transplantation tumors in vivo, and prolongs the survival time of tumor-bearing mice. The present invention further provides a monoclonal antibody production cell line mouse hybridoma cell line HC-7B2 (with the preservation number of CGMCC No.10897).

This invention provides an improved process for manufacturing a Rabies monoclonal antibody (HuMab 17C7) that results in low osmolality, minimum secondary metabolites like ammonia and lactate, enhanced cell growth and productivity, minimum aggregation or degradation of monoclonal antibody during purification, thereby improving potency of monoclonal antibody (HuMab 17C7) as compared to human rabies immunoglobulin (hRIG).

The present invention discloses an anti-human-lung-cancer-stem-cell monoclonal antibody, and particularly relates to a new monoclonal antibody capable of specifically recognizing and inhibiting human lung cancer stem cells, a hybridoma cell line for producing the monoclonal antibody, and applications of the monoclonal antibody in preparation of drugs for treatment of human lung cancer recurrence. According to the present invention, the monoclonal antibody can significantly inhibit the self-renewal, the invasion and the migration of human lung cancer stem cells in vitro, can significantly inhibit the growth and the recurrence of human lung cancer transplantation tumors in vivo, and prolongs the survival time of tumor-bearing mice. The present invention further provides a monoclonal antibody production cell line mouse hybridoma cell line LC-5A6 having the preservation number of CGMCC No.10898.

The invention relates to a preparation method of an alicyclic acid bacillus resisting monoclonal antibody. The method comprises the following steps: (1) preparing an immunized mouse; (2) culturing myeloma cells for 3-4 days to obtain cultured mouse myeloma cells; (3) inoculating alicyclic acid bacillus in the abdominal cavity of the immunized mouse for immunization and obtaining spleen cells; dispersing the spleen cells to obtain a spleen cell precipitate; (4) centrifuging the cultured mouse myeloma cells to obtain a myeloma cell precipitate; (5) respectively suspending and mixing the spleen cell precipitate and the myeloma cell precipitate in a DMEM high-sugar culture solution, and centrifuging to obtain a precipitate; centrifuging and suspending the precipitate to obtain fused cells; (7) culturing and assaying the fused cells to determine hybridoma cells; cloning and screening the hybridoma cells to obtain a hybridoma cell line; and (8) preparing a monoclonal antibody, namely performing monoclonal antibody production on the hybridoma cell line according to a general method. The preparation method disclosed by the invention is rapid and high in specificity.

The present invention relates to a preparation method of a regenerated gene protein REG1 alpha monoclonal antibody. The method comprises the following steps: (1) preparing immunized mice; (2) culturing myeloma cells for 3-4 days to obtain cultured mouse myeloma cells; (3) dissolving REG1 alpha protein, inoculating abdominal cavity of the immunized mice with the dissolved REG1 alpha protein for immunization, obtaining splenocytes, and dispersing the splenocytes to obtain a splenocyte precipitate; (4) centrifuging the cultured mouse myeloma cells to obtain a myeloma cell precipitate; (5) respectively suspending the splenocyte precipitate and the myeloma cell precipitate by using a DMEM high-glucose culture solution, conducting mixing, and centrifuging the mixture to obtain a precipitate; (6)centrifuging and suspending the precipitate to obtain fusion cells; (7) culturing and determining the fusion cells, and determining hybridoma cells; (8) screening and separating monoclonal antibodies; and (9) preparing a large amount of monoclonal antibodies, namely producing the monoclonal antibodies from a hybridoma cell line according to a conventional method. The preparation method is rapid and high in specificity.

The present invention discloses a bioreactor cell culture apparatus capable of onlinely and continuously monitoring physiological and biochemical indexes in a variety of cell culture environments and directly regulating and controlling cell states (including cell cycle and the like), and a method for producing anti-Her-2 monoclonal antibodies through the apparatus. With the apparatus and the method of the present invention, the bacterial pollution risk during the cell culture process can be reduced, the stable cell growth environment can be maintained, the constant cell growth viability can be maintained, and the high-quality, high-efficiency and continuous anti-Her-2 monoclonal antibody production can be achieved.

The invention belongs to the technical field of cell engineering and discloses a unicellular fusion method for pig and mouse cells. The unicellular fusion method is provided aiming at the problem of current low xenogenic cell fusion efficiency. The method includes: taking mouse embryonic stem cells and pig pluripotent stem cells or pig fibroblast cells as cellular fusion objects; adopting digestive juice containing pancreatin in mass fraction of 0.25% to digest the pig and mouse cells into single cells, culturing, and adopting cell agglutinin for constructing one-to-one unicellular pairs; culturing for 1min in polyethylene glycol in mass fraction of 50%, and continuing cell culture for seven days to obtain fused cells of pig and mouse single cells. The unicellular fusion method is applicable to animal breeding research or monoclonal antibody production.

The invention discloses an affinity purification method for reducing the content of host cell protein in production of an anti-human interleukin-33 monoclonal antibody, which comprises the following steps: balancing before loading: balancing an affinity chromatography medium by using a buffer solution I to obtain the balanced affinity chromatography medium; loading a sample: combining the anti-human interleukin-33 monoclonal antibody fermentation liquor with the balanced affinity chromatography medium; and elution: carrying out pre-elution and final elution on the mixed solution so as to remove host cell protein and obtain the purified anti-human interleukin-33 monoclonal antibody, wherein the anti-human interleukin-33 monoclonal antibody comprises three heavy chain complementarity determining regions and three light chain complementarity determining regions, the three heavy chain complementarity determining regions are CDR-H1, CDR-H2 and CDR-H3, and the three light chain complementarity determining regions are CDR-L1, CDR-L2 and CDR-L3. The method is simple and easy to implement, amplification purification production can be carried out, the cell fermentation supernate does not need to be pretreated, the elution sample yield is high, the HCP residual quantity is kept at a low level, and therefore the pressure for removing HCP in the subsequent purification step is relieved.

The invention discloses a single antigen specific transgenic hybridoma cell screening method, which comprises the following steps: preparing a transgenic Sp2/0 myeloma cell expressing EGF-R-AVI-tag fusion protein, and combining with a splenocyte fusion technology of a traditional hybridoma technology to obtain a transgenic hybridoma cell capable of displaying a secreting type specific antibody on a cell membrane. And screening of a single specific transgenic hybridoma cell is realized by using a single cell sorting system. The invention provides an accurate and high-throughput transgenic hybridoma monoclonal antibody production platform based on a transgenic technology. Compared with the traditional hybridoma technology, the preparation time and cost of a single antigen-specific hybridoma cell are remarkably shortened, the single cell rate and positive rate are improved, and the titer and affinity of the prepared monoclonal antibody are also remarkably improved. The transgenic hybridoma antibody preparation technology provided by the invention can be used for preparing rare monoclonal antibodies with high specificity, high sensitivity and high stability, and has a wide application prospect.

The invention relates to molecular biology and particularly discloses a gene sequence capable of preventing gradual inactivation of a promoter in subculture and applications of the gene sequence. The nucleotide sequence of the gene sequence is shown as SEQ ID No.1. The gene sequence is used for constructing a carrier and a CHO cell line, can prevent gradual inactivation of the promoter in subculture of CHO cells and can be used for monoclonal antibody production through the CHO cell line to increase the monoclonal antibody yield.