Serum-free culture composition and its application
A technology of serum-free culture and serum-free medium, which is applied in the direction of culture process, tissue culture, cell culture active agent, etc., can solve the problem of large fluctuations in the expression of monoclonal antibodies, low expression of monoclonal antibodies, time-consuming and labor-intensive problems, etc. question
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[0046] The method for preparing a hybridoma cell-based monoclonal antibody according to one embodiment is characterized in that it includes the following operations S110-S120:
[0047] S110. Directly inoculate the hybridoma cells into the above-mentioned serum-free culture composition for culture, separate the solid from the liquid, and obtain a supernatant.
[0048] In one embodiment, the hybridoma cells are mouse hybridoma cells. It should be noted that the hybridoma cells are not limited to mouse hybridoma cells, and may also be rat hybridoma cells.
[0049] Further, the hybridoma cells are hybridoma cells capable of producing CA242 antibody, hybridoma cells capable of producing CY211 antibody or hybridoma cells capable of producing CKMB antibody.
[0050] Wherein, the CA242 antibody is an antibody capable of specifically recognizing CA242. CA242 is a sialylated carbohydrate antigen, which is clinically used in the diagnosis of digestive tract malignant tumors, especially...
Embodiment 1
[0066] There were 11 groups in the experiment, numbered A to K, and the cell lines were all cell line 1, which secreted CA242 antibody. The culture process of each group of cell lines is as follows:
[0067] (1) Prepare serum-free medium according to the instructions, and add EGF, FGF, albumin and hydrolyzate to the serum-free medium to obtain serum-free culture composition; the addition amount of EGF, FGF, albumin and hydrolyzate is as shown in the table 1, wherein the hydrolyzate used in groups A to K is yeast hydrolyzate.
[0068] (2) Put the serum-free culture composition into the shake flask after aseptic filtration, inoculate the unaccustomed cell line 1 into the serum-free culture composition at a density of 3×10^5 cells / mL, and culture in a shaker ; culture conditions: 8% CO 2 , the rotational speed is 120rpm, the amplitude is 50mm, and the temperature is 37°C. Samples were taken every 24 hours to detect the density of viable cells and the expression of monoclonal a...
Embodiment 2
[0086] There were 6 groups in the experiment, numbered a to g, and the cell lines were all cell line 2, which secreted CY211 antibody. The details of the culture process of each group of cells are as follows:
[0087] (1) Prepare serum-free medium according to the instructions, and add EGF, FGF, albumin and hydrolyzate to the serum-free medium to obtain serum-free culture composition; the addition amount of EGF, FGF, albumin and hydrolyzate is as shown in the table 2. Wherein, the hydrolyzate used in groups a to d is yeast hydrolyzate. The hydrolyzate used in group e is hydrolyzed soybean protein. The hydrolyzate used in group f is a mixture of yeast hydrolyzate and hydrolyzed soybean protein, and the mass ratio of yeast hydrolyzate to hydrolyzed soybean protein is 1:1. The hydrolyzate used in group g is a mixture of yeast hydrolyzate and hydrolyzed vegetable protein, and the mass ratio of yeast hydrolyzate to hydrolyzed vegetable protein is 1:1, and hydrolyzed vegetable pr...
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