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Falciparum malaria multi-epitope vaccine M.RCAg-1 protein and M.RCAg-3 compatibility studies of different adjuvants

Author: MaRuiSen
Tutor: WangHeng
School: Beijing Union Medical College
Course: Pathogen Biology
Keywords: Plasmodium falciparum Multi-epitope vaccine Adjuvant compatibility
CLC: R392
Type: Master's thesis
Year: 2011
Downloads: 24
Quote: 1
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Abstract


Malaria is one of the world's three major communicable diseases. According to the latest WHO statistics show that in 2009 225 million people worldwide infected with malaria, there are 78.1 million people in his death, and most are children under five and pregnant women [1]. In all Plasmodium falciparum, Plasmodium falciparum is the most serious hazards, one of the highest death rate. In malaria control in the process emerging drug resistance in Plasmodium falciparum and Anopheles, which encourages people to preventive and curative malaria vaccine needs. Because the life cycle of the malaria parasite complex, stage-specific antigens, insect strains differences and highly variable characteristics, present for a single antigen or epitope vaccine in the relevant experiments have not obtained good results, so multi-epitope chimeric vaccine against malaria vaccine research has become a hot spot [2]. Our previous work, we use a random epitope recombinant technology to build multi-epitope gene pool [3] and serum screening with libraries optimal immunogenicity, stability, random combination of the best artificial antigen M.RCAg- 1 and M.RCAg-3. Freund's adjuvant in mice and rabbits both animal models have demonstrated strong immunogenicity and in vitro inhibition of the growth of P. falciparum (Growth Inhibi tion Assay.GIA) by switching adjuvant Montanide ISA51 (Seppic, France) immune rhesus found greater local irritation. Therefore, the purpose of this study is to select new antigens and adjuvants that M.RCAg-1 M.RCAg-3 can successfully be used in clinical observation in this study on the basis of previous work, based on the second generation of multi-epitope chimeric antigen vaccine M.RCAg-3 physicochemical properties, the use of an anion exchange column, hydrophobic interaction chromatography and molecular sieve, explored and summed up the second generation of multi-epitope chimeric antigen vaccine M.RCAg-3 purification steps. And selection of a clinical application or adjuvants in clinical applications (CpG.Al.AD.ADL) and M.RCAg-1 and compatibility M.RCAg-3, to increase its immunogenicity. Also used nanoparticles wrapped protein or protein adjuvant to immunize animals to explore new antigen presenting ways to increase the immunogenicity of the antigen. Through the evaluation of the effectiveness of different groups immune to analyze the corresponding adjuvant effects. The final choice ADL and AD respectively M.RCAg-1 and M.RCAg-3 compatibility immune rabbits, verify its antibody in vitro protective effect. Mainly achieved the following milestones: 1. Worked out the second-generation multi-epitope purified chimeric antigen vaccine M.RCAg-3 purification steps, the final purification result can be achieved 98.8%. 2 Screening and M.RCAg-1 and M.RCAg-3-phase compatible adjuvants. ADL and AD respectively adjuvant can significantly enhance M.RCAg-1 and M.RCAg-3 immune response. The adjuvant CpG and CpG A1 enhanced antigen immune response capacity is relatively weak. Initially identified using nano-wrapped presenting proteins method is not suitable M.RCAg-1 and M.RCAg-3. 3. M. RCAg-1 (small scale), M.RCAg-1 (pilot) and M.RCAg- 3, respectively, with adjuvant ADL AD1 and AD compatibility immune rabbits, the total IgG and specific IgG levels close to the WHO requirements GIA positive control antibody in vitro growth inhibition assay (GIA) standards.

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