A method for producing recombinant canine parvovirus monoclonal antibody

Abstract

The present invention relates to the technical field of cell recombinant antibody, in particular to a medium for SP2 / 0 cell perfusion and a method for producing recombinant canine parvovirus monoclonal antibody, including medium preparation, tumor cell domestication, fed-batch culture and antibody purification; wherein The culture device includes a reaction tank, a replenishment tank and a controller; during the process of feeding batches, the fed-in medium components cannot be fully replenished to the area consumed by the proliferation of recombinant tumor cells, which weakens the nutrients in the fed-in medium Therefore, the present invention makes the rotating and shaken culture medium components in the supplement tank move along the reaction tank under the action of extra kinetic energy by setting the groove around the inner wall of the reaction tank. The inner wall of the recombinant tumor cell is washed into the culture medium of the recombinant tumor cell, so that the supplemented culture medium is evenly distributed among the recombinant tumor cells, thereby improving the application effect of the production method of the recombinant canine parvovirus monoclonal antibody.

Description

technical field [0001] The invention relates to the technical field of cell recombinant antibody, in particular to a production method of recombinant canine parvovirus monoclonal antibody. Background technique [0002] With the substantial increase of working dogs and pet dogs in people's daily life, the life and health of dogs has gradually attracted attention, and canine parvovirus is one of the most serious infectious diseases that endanger dogs. Serum and monoclonal antibody for treatment; in vitro culture of canine parvovirus monoclonal antibody has become an effective means of clinical diagnosis and treatment; SP2 / 0 cells are myeloma cells, and their in vitro unlimited proliferation characteristics are used for the development of recombinant antibodies. However, the expression level of monoclonal antibodies cultured in vitro is at a low level, which hinders the promotion of canine parvovirus monoclonal antibody therapy; for the introduction of recombinant monoclonal an...

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Problems solved by technology

[0003] In order to increase the expression of monoclonal antibodies in vitro, the optimization of the culture medium and the design of the culture process of recombinant myeloma cells become the key; dozens of components are added to the emerging serum-free medium to meet the proliferation of tumor cells and avoid serum components. It brings the risk of botulinum toxin in the proliferation of tumor cells, as well as the difficulty of subsequent separation and purification, but its tedious optimization design brings limited improvement in the expression of monoclonal antibodies, and at the same time increases the difficulty of continuous supplementation of the perfusion medium

[0004] In the prior art, there are also some technical solutions about the medium for SP2 / 0 cell perfusion and the production method of recombinant monoclonal antibody. For example, a Chinese patent with application number 2009100664783 discloses a hybridoma cell for preparing monoclonal antibody Culture medium for cloning and its preparation method, prepare RPMI-1640 complete medium according to the conventional method, and then mix it with a certain proportion of cell-free culture supernatant of BHK-21, CRFK, Hela and other cells, complete medium and each cell The volume ratio of the supernatant is 7:1-1:1; the medium for hybridoma cell cloning provided by this technical solution can completely omit the use of BALB / c mouse peritoneal macrophages as the medium in the conventional hybridoma cell cloning process. The link of feeder cells greatly simplifies the cloning operation and significantly increases the growth rate of the cloned cells; however, this technical solution does not solve the problem of the difference between the hybridoma cells in the medium perfused in batches during the proliferation of hybridoma cells. The problem of uneven distribution reduces the survival rate of tumor cells, thereby weakening the effective expression of monoclonal antibodies in tumor cells

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