Pilot Scale Production and Identification of Plasmodium Falciparum Multiepitope Random Constructed Vaccine M.RCAg-1
1 Institute of Basic Medical Sciences，Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing 100005, China
2 Beijing Weishibohui Chromatography Technology Co., Ltd, Beijing 100176, China
Abstract：The aim of this work is to produce Plasmodium falciparum multiepitope random constructed vaccine M.RCAg-1 efficiently in pilot scale and identify its immunogenicity. M.RCAg-1 producing E.coli was fermented in 20 L culture. After the fermentation products were homogenized by high pressure, the obtained bacteria lysate was purified by Ni FF affinity chromatography and G200 HP gel filtration chromatography. Fifteen BALB/c mice were randomly divided into three groups (the group of Freund’s adjuvant, the group of pilotscale M.RCAg-1 combined with Freund’s adjuvant and the group of labscale M.RCAg1 combined with Freund’s adjuvant) and injected subcutaneously. The serum of these mice was collected after third immunization to analyze the specific serum antibody by indirect ELISA and indirect immunofluoreseence assay (IFA) technology. Fermentation process was finished after 4 h induced by IPTG. The collected wet cell yield was 25 g/L, and the expression level of M.RCAg-1 was about 24%. After twostep purification, the final M.RCAg-1 was over 95% in purity，the yield was 53.2%．Compared with the control group, both the pilotscale and labscale M.RCAg-1 induced high level antibody response in mice and the antibody titter was 1∶1 024 000.At the same time, serum antibody of the two experimental groups recognized the natural antigen of Plasmodium falciparum very well and there was no difference of recognition between the two groups(P>0.05). With the successful production of Plasmodium falciparum multiepitope random constructed vaccine M.RCAg-1 efficiently in pilot scale and the confirmation of its good immunogenicity in mouse model, this study provids the foundation for preclinic study of M.RCAg-1.
刘新1 韦新桂2 &n. 恶性疟原虫多表位人工随机重组疫苗M.RCAg-1的中试规模制备与鉴定[J]. 中国生物医学工程学报, 2014, 33(3): 329-334.
LIU Xin1 WEI XinGui2 GAO Yu\|Hui1 WANG Heng1*. Pilot Scale Production and Identification of Plasmodium Falciparum Multiepitope Random Constructed Vaccine M.RCAg-1. journal1, 2014, 33(3): 329-334.
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