Optimized Purification of CIDRα-PfEMP1 Plasmodium falciparum Recombinant Protein with Affinity Chromatography

  • Eqiel Navadz Akthar Alami Fakultas Kedokteran, Universitas Jember
  • Erma Sulistyaningsih Fakultas Kedokteran, Universitas Jember; Center for Excellence of Agromedicine (CEAMED), Universitas Jember
  • Irawan Fajar Kusuma Fakultas Kedokteran, Universitas Jember
  • Sheilla Rachmania Fakultas Kedokteran, Universitas Jember


Interaction of Cysteine-rich Interdomain Region (CIDR)α-Plasmodium falciparum Erythrocyte Membrane Protein (PfEMP1) and Endothelial Cell Receptors especially CD36 on host cells is main malaria pathogenesis, makes this domain as a malaria vaccine candidate. Recently, the development of the malaria vaccine is conducted by recombinant technology, and the purification of the CIDRα-PfEMP1 recombinant protein is a pivotal step. This study aimed to determine an optimal condition to purify the CIDRα-PfEMP1 recombinant protein by affinity chromatography through imidazole and NaCl concentration. The purified recombinant protein was visualized using SDS-PAGE and its concentration was measured using Image J software and Bradford Assay. The data were analyzed using SPSS 26 software, and the Paired T-Test analysis was conducted to compare the concentration of purified recombinant protein from two different methods. The result showed that thetarget band of purified recombinant protein was 27 kDa. The thickest target protein band was observed in purified recombinant protein using 140 mM imidazole and 300 mM NaCl. The recombinant protein concentration using Image J software was 0.025 µg/µL, while the Bradford Assay was 0.56 µg/µL. The Paired T-Test analysis has a significance value of 0.010 (p<0.05), meaning there was a significant difference between the concentration measurement using Image J software and Bradford Assay. In conclusion, the optimized condition to purify the CIDRα-PfEMP1 recombinant protein by affinity chromatography was using 140 mM imidazole and 300 mM NaCl. It is suggested to measure the purified CIDRα-PfEMP1 recombinant protein concentration using the Bradford Assay method due to its convenience and sensitivity.


Dahlan SM. 2014. Statistik untuk Kedokteran dan kesehatan: Deskriptif, Bivaruat, dan Multivariat. (W. Kurniawan (ed.). 6th ed., Issue 1). Salemba Medika.

Dewi R, Ratnadewi AAI, Sawitri WD, Rachmania S & Sulistyaningsih E. 2018. Cloning, Sequence Analysis, and Expression of CIDR1α-pfEMP1 from Indonesian Plasmodium falciparum isolate. Current Topics in Peptide and Protein Research. 19:95-104.

Ernst O & Zor T. 2010. Linearization of the Bradford protein assay. Journal of Visualized Experiments. 38:1-6.

Ferreira T & Rasband W. 2012. Image J User Guide User Guide ImageJ. Image J User Guide, 1.46r.

Flick K, Ahuja S, Chene A, Bejarano MT & Chen Q. 2004. Optimized Expression of Plasmodium falciparum Erythrocyte Membrane Protein 1 Domains in Escherichia coli. Malaria Journal. 3(50):1-8.

Gräslund S, Nordlund P & Weigelt J. 2008. Protein Production and Purification. Nature Methods. 5(2):135-146.

IBA Lifesciences. 2012. Expression and Purification of Proteins Using 6x Histidine -tag A Comprehensive Manual. In IBA Solutions For Life Sciences (Issue October). IBA Headquarters.

Lubiana P, Bouws P, Roth LK, Dörpinghaus M, Rehn T, Brehmer J, Wichers JS, Bachmann A., Höhn K, Roeder T, Thye T, Gutsmann T, Burmester T, Bruchhaus I & Metwally NG. 2020. Adhesion Between P. falciparum Infected Erythrocytes and Human Endothelial Receptors Follows Alternative Binding Dynamics under Flow and Febrile Conditions. Scientific Reports. 10(1):1-14.

Oliveira C & Domingues L. 2018. Guidelines to Reach High-quality Purified Recombinant Proteins. Applied Microbiology and Biotechnology. 102(1): 81-92.

Olson BJSC & Markwell J. 2007. Assays for Determination of Protein Concentration. in Current Protocols in Protein Science (pp. 1-29). John Wiley & Sons Inc.

Puspitaningrum R, Adhiyanti C & Solihin. 2018. Genetika Molekuler dan Aplikasinya (S. Attas (ed.)). Deepublish.

Putri M, Munadziroh E & Soebagio. 2015. Purifikasi Secretory Leukocyte Protease Inhibitor Membran Amnion Rekombinan sebagai Kandidat Biomaterial Menggunakan Kromatografi Afinitas. Jurnal PDGI. 64(1): 11-16.

Qiagen. 2003. The QIA Expressionist - Fifth Edition. In Qiagen GmbH, Düsseldorf. Germany (Issue June).
Reinking L. 2007. ImageJ Basics. Word Journal of The International Linguistic Association. 1-22.

Riskesdas K. 2018. Hasil Utama Riset Kesehatan Dasar (RISKESDAS). Journal of Physics A: Mathematical and Theoretical. 44(8):1-200.

Sawitri WD, Narita H, Ishizaka-Ikeda E, Sugiharto B, Hase T & Nakagawa A. 2016. Purification and Characterization of Recombinant Sugarcane Sucrose Phosphate Synthase Expressed in E. Coli and Insect Sf9 cells: An Importance of the N-Terminal Domain for An Allosteric Regulatory property. Journal of Biochemistry. 159(6): 599-607.

Simaremare APR, Bela B, Yasmon A & Ibrahim F. 2015. Optimizations of Expression and Purification of Recombinant HIV-1 CRF01_AE p24 Protein in Escherichia coli for Development of Immunodiagnostic
Assay. Medical Journal of Indonesia. 24(1):14-18.

Smith JD. 2014. The Role of PfEMP1 Adhesion Domain Classification in Plasmodium falciparum Pathogenesis Research. Mol Biochem Parasitol. 195(2): 82-87.

Sulistyaningsih E, Armiyanti Y & Dewi R. 2021. The Cidr1α-Pfemp1 Sequence From Indonesian Plasmodium falciparum and Its Potential Association with The Cerebral Outcome. MNJ (Malang Neurology Journal). 7(1):34-39.

Sulistyaningsih E, Fitri LE, Löscher T & Berens-Riha N. 2013. Diversity of The Var Gene Family of Indonesian Plasmodium falciparum Isolates. Malaria Journal. 12(1): 1-13.

Walker JM. 2002. The Protein Protocols Handbook-Second Edition. Humana Press Inc.

World Health Organization. 2019. World malaria report 2019. In WHO Regional Office for Africa. World Health Organization.

Zhang CL, Zhou HN, Liu Q & Yang YM. 2019. Genetic Polymorphism of Merozoite Surface Proteins 1 And 2 of Plasmodium Falciparum in The China-Myanmar Border Region. Malaria Journal. 18(1):1-10.
How to Cite
ALAMI, Eqiel Navadz Akthar et al. Optimized Purification of CIDRα-PfEMP1 Plasmodium falciparum Recombinant Protein with Affinity Chromatography. Jurnal ILMU DASAR, [S.l.], v. 23, n. 2, p. 107-112, july 2022. ISSN 2442-5613. Available at: <https://jurnal.unej.ac.id/index.php/JID/article/view/24127>. Date accessed: 01 mar. 2024. doi: https://doi.org/10.19184/jid.v23i2.24127.