Expression and purification of Plasmodium falciparum MSP-1(42): A malaria vaccine candidate

J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Mar 25;786(1-2):61-72. doi: 10.1016/s1570-0232(02)00722-5.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chromatography, Affinity
  • Cloning, Molecular
  • Cross Reactions
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Malaria Vaccines / genetics*
  • Malaria Vaccines / isolation & purification*
  • Merozoite Surface Protein 1 / chemistry*
  • Molecular Sequence Data
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology*
  • Peptide Fragments / isolation & purification
  • Plasmodium falciparum / immunology*
  • Sequence Homology, Amino Acid

Substances

  • Malaria Vaccines
  • Merozoite Surface Protein 1
  • Peptide Fragments