New use of co-stimulation factor and fusion protein
A fusion protein and application technology, applied in the direction of peptide/protein components, medical preparations containing active ingredients, drug combinations, etc., to achieve significant therapeutic activity
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Embodiment 1
[0028] Example 1 Preparation of B7.1-Fc in the present invention
[0029] The B7.1-Fc fusion protein was prepared by gene recombination technology. According to the method well known to those skilled in the biological field, the B7.1-Fc expression vector was obtained and constructed by PCR technology, and transferred to mouse myeloma NSO cells to screen to obtain long-term stable expression of the fusion protein B7 .1-Fc engineered cells, then through fed-batch culture to obtain high-expressing B7.1-Fc culture supernatant, and then purified by protein A affinity chromatography and ion exchange chromatography to obtain high-purity fusion protein, 4°C save.
[0030] The preparation method of B7.1-Fc in the present invention can refer to the preparation method in CN200580042109.0. The B7.1-Fc refers to B7.1 / IgG1Fc, that is, the fusion protein of B7.1 and IgG1Fc. See the sequence table Aihong Liu, Peisheng Hu; Clin Cancer Res 2005; 11(23) December 1, 2005, 8492-8502.
[0031] Determine...
Embodiment 2
[0032] Example 2 Pharmacodynamic experiment of immune costimulatory factor fusion protein B7.1-Fc
[0033] 1. Test sample and dose design
[0034] Name: Immune costimulatory factor fusion protein B7.1 / IgG1Fc (abbreviated as B7.1-Fc) was prepared according to Example 1
[0035] Provider: Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences
[0036] Properties: colorless, clear liquid
[0037] Storage conditions: 4℃ storage
[0038] Administration route: tail vein injection
[0039] Solvent: physiological saline
[0040] 2. Positive control:
[0041] Cyclophosphamide for Injection; Paclitaxel
[0042] 3. Tumor cell lines:
[0043] Mouse cervical cancer U14, mouse sarcoma S180, mouse melanoma B16-F10, all can be obtained by purchasing commercially available products
[0044] 4. Animals:
[0045] BALB / c mice, SPF / VAF grade provided by Beijing Huafukang Biotechnology Co., Ltd., certificate number SCXK (京)2009-0004
[0046] 5. Conditions of breeding facilities:
[0047] Barr...
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