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
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.
何学虎 蔺亚晖 王恒*. 恶性疟原虫多表位疫苗M.RCAg-1在大肠杆菌中的表达和纯化[J]. 中国生物医学工程学报, 2012, 31(2): 285-290.
E Xue Hu LIN Ya Hui WANG Heng*. Expression and Purification of Plasmodium Falciparum Multiepitope Chimeric Vaccine M.RCAg-1 in Escherichia Coli. journal1, 2012, 31(2): 285-290.
