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Expression Of Murine IFN-? And IL-10 Cytokines And Preparation Of Monoclonal Antibodies

Posted on:2021-05-24Degree:MasterType:Thesis
Country:ChinaCandidate:T JiangFull Text:PDF
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Cytokines are a class of small-molecule soluble proteins that play a very important role in the innate and adaptive immunity of animals.As more important cytokines,IFN-? and IL-10 have attracted the attention of many researchers.IFN-? and IL-10 are the two most potent cytokines with the most extensive systemic immune response.IFN-? can directly mobilize the proliferation and differentiation of various immunogenic cells in the body and participate in the immune response of all type I helper T cells(Th1 cells),IL-10 can inhibit the inflammatory response and improve the body's immune system to foreign bacterial and Viral response.IFN-? and IL-10 are closely related to many diseases in animals.This experiment first used genetic engineering to express and purify murine IFN-? and IL-10 proteins.Using monoclonal antibody technology,15 hybridoma cells stably secreting antibodies(two hybridoma cell lines secreting IFN-? antibody and thirteen hybridoma cell lines secreting IL-10 antibody)were screened.The subtype identification of the hybridoma cell line 2D8 secreting IFN-? antibody was Ig G2 a.Hybridoma cells 2D8 were injected into the abdominal cavity of mice treated with Freund's incomplete adjuvant at a dose of 106 each,and IFN-? antibody was obtained by ascites collection and purification.The IFN-? and IL-10 proteins expressed in this work can provide an important material basis for other researchers to further study the structure and function of mice and the relationship with related diseases.The hybridoma cells obtained by monoclonal antibody technology can be the subsequent establishment of a double-antibody sandwich ELISA method to detect these two cytokines provides critical antibody support.
Keywords/Search Tags:IFN-?, IL-10, cell fusion, monoclonal antibody, Hybridoma cell
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