Expression and purification of Plasmodium falciparum MSP-142:: A malaria vaccine candidate
Expression and purification of Plasmodium falciparum MSP-142:: A malaria vaccine candidate
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DOI：
10.1016/s1570-0232(02)00722-5
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发表时间：
2003-03-25
影响因子：
3
通讯作者：
Lutz, R
中科院分区：
文献类型：
--
作者：
Epp, C;Kauth, CW;Lutz, R
The C-terminal 42.10(3) Da portion of the merozoite surface protein (MSP-1) of the human malaria parasite Plasmodium falciparum is of interest, not only because it may constitute an essential part of a future anti-malaria vaccine, but also due to its role during the infection of erythrocytes by the parasite. We have cloned and expressed two synthetic DNA sequences encoding the two prototypic MSP-1(42) variants in E. coli. When over-produced, both proteins form insoluble aggregates which were isolated in high purity and yield. After solubilisation and refolding in vitro, both proteins were purified to homogeneity by a three-step procedure applying Ni-chelate, size exclusion and immuno-affinity chromatography. After purification, both proteins meet key criteria of preparations for clinical use. First, conformational studies suggest proper folding of the proteins, particularly in the region containing two EGF-like domains. Polyclonal serum raised against E. coli produced MSP-1(42) recognizes native MSP-1 in Plasmodium infected erythrocytes as shown by immunofluorescence. (C) 2002 Elsevier Science B.V. All rights reserved.
