Preparation method of tumor dendritic cell vaccine sensitized by glycosylated MUC1 (mucoprotein 1) antigen
A technology for vaccine preparation and glycosylation, which is applied in the field of preparation of anti-tumor immune cell vaccines, and can solve the problem of low efficiency of antigen presentation.
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[0022] The specific technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and embodiments, so that those skilled in the art can understand the present invention without restricting their rights.
[0023] 1. Preparation of MUC1M-FP (such as figure 1 Shown):
[0024] 1) Preparation of MUC1FP: A 309 base fragment (pDF9.3, encoding a 60 base motif fragment in the VNTR region of MUC1 cDNA slightly more than 5) was cloned into the bacterial expression vector pGEX-3X for correct expression Frame and express direction. MUC1-FP (including GST and MUC1VNTR) was induced with IPTG. The cells were collected by centrifugation, washed and sonicated (in a buffer containing TritonX-100), the supernatant containing the solubilized protein MUC1-FP was mixed with glutathione-coated magnetic beads (disulfide bond connection), and then collected by centrifugation . The fusion protein MUC1-FP was washed with a buffer containing reduc...
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