Purification of NTS-DBL1α Malaria Protein

E. coli codon optimized NTS-DBL1α^It4var60 domain of the parasite clone FCR3S1.2 were expressed as a C-terminal 6x histidine tagged recombinant proteins in the E. coli strain BL21 DE3 (ΔslyD) as described in [12] (link). Bacteria cultures were grown to OD₆₀₀ = 0.8, then induced for 3 h at 37°C with 0.1 mM IPTG. Proteins were purified by Immobilized Metal Affinity Chromatography over Ni-NTA columns (Qiagen), eluted with 500 mM imidazole and further purified to homogeneity by size exclusion chromatography on a HiLoad 16/60 Superdex 75 pg colum (GE-Healthcare) [12] (link).

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Albrecht L., Angeletti D., Moll K., Blomqvist K., Valentini D., D'Alexandri F.L., Maurer M, & Wahlgren M. (2014). B-Cell Epitopes in NTS-DBL1α of PfEMP1 Recognized by Human Antibodies in Rosetting Plasmodium falciparum. PLoS ONE, 9(12), e113248.

Publication 2014

Ni-NTA columns

Affinity chromatography Bacteria Clone Codon E coli E coli proteins Histidine Imidazole Iptg Metal Parasite Proteins Size exclusion chromatography Strain

Corresponding Organization : Karolinska Institutet

Other organizations : Karolinska University Hospital, Universidade Estadual de Campinas (UNICAMP)

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Variable analysis

independent variables

IPTG concentration (0.1 mM)

dependent variables

Purified recombinant protein levels

control variables

E. coli strain BL21 DE3 (ΔslyD)
Bacterial culture growth to OD600 = 0.8
3 h induction time at 37°C

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