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| Expression and Purification of Plasmodium Falciparum Multiepitope Chimeric Vaccine M.RCAg-1 in Escherichia Coli |
| Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing 100005, China |
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Abstract This study is aimed to express and purify Plasmodium falciparum multiepitope chimeric vaccine M.RCAg-1 in Escherichia coli and to maximize the purity and yield of target protein. Extraction and sequence of the recombinant plasmid from the engineering bacteria BL21 (DE3)M.RCAg-1/pDS-eX, IPTG was used to induce to express M.RCAg-1 Five different methods of combinations of Ni affinity purification, anion exchange chromatography and gel filtration chromatography were used to purify M.RCAg-1, and the optimal purification approach was selected according to the final purity and yield of M.RCAg-1. The specificity of Ni-NTA Agarose was much better, by Ni-NTA Agarose first step purification, the M.RCAg-1 purity could reach 79.98%, the next two-step anion exchange chromatography and gel filtration could enhance M.RCAg-1 purity significantly and the final M.RCAg-1 was over 99% in purity, yield was above 15%. This research has established a protocol for expressing and purifying Plasmodium falciparum multiepitope chimeric vaccine M.RCAg-1 in Escherichia coli and formed the foundation for the downstream large scale production and further investigates the M.RCAg-1 effectiveness.
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