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| Construction,Expression,Purification of VEGFⅡ/GRP Fusion Protein and the Effects on RM-1 Prostate Tumor in Mice |
| CAO Rong-yue, YU Min-xia, ZHANG Xin-li, LI Man-man, MIAO Zi-tao, JIN Liang |
| School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China |
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Abstract An expression vector pET-28a-VEGFⅠ-M2-GRP plasmid containing the VEGF and GRP antigen epitopes was designed and constructed. After being expressed in recombined strains induced by lactose, the fusion protein was lysised by ultrasonic. The resulting inclusion body was washed,dissolved, and dialysised for renaturation and purified by ion-exchange chromatograph to get the target protein.VEGFⅡ/GRP was identified by Western blot. The C57BL/6J mice subcutaneous tumor model of prostate cancer cell line RM-1was established to receive treatments with VEGFⅡ/GRP vaccine.The tumor growth,the blood vessel densities and the concentration of anti-VEGF antibody were examined to assess the effect of anti-angiogenesis and anti-tumor. ELISA showed that the concentration of anti-VEGF antibody in the mice serum of VG group was significantly higher than that of NS group (P<0.05),the anti-angiogenesis effect of VG group was superior to that of NS group (P<0.05).Thus,the constructed fusion protein can inhibit angiogenesis.
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Received: 01 February 2016
Published: 25 August 2016
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