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

doi:10.3969/j.issn.0258-8021. 2014. 03.010

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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 pilotscale M.RCAg-1 combined with Freund’s adjuvant and the group of labscale M.RCAg1 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 twostep purification, the final M.RCAg-1 was over 95% in purity，the yield was 53.2%．Compared with the control group, both the pilotscale and labscale 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 preclinic study of M.RCAg-1.

Key words： plasmodium falciparum protein vaccine pilot scale fermentation immunogenicity

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	LIU Xin¹WEI XinGui²GAO Yu\\|Hui¹WANG Heng^1*

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LIU Xin¹WEI XinGui²GAO Yu\|Hui¹WANG Heng^1*. Pilot Scale Production and Identification of Plasmodium Falciparum Multiepitope Random Constructed Vaccine M.RCAg-1[J]. journal1, 2014, 33(3): 329-334.

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http://cjbme.csbme.org/EN/10.3969/j.issn.0258-8021. 2014. 03.010 OR http://cjbme.csbme.org/EN/Y2014/V33/I3/329

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