Application of crossflow ultrafiltration for scaling up the purification of a recombinant ferritin

Federica Palombarini; Francesca Ghirga; Alberto Boffi; Alberto Macone; Alessandra Bonamore

doi:10.1016/j.pep.2019.105451

Application of crossflow ultrafiltration for scaling up the purification of a recombinant ferritin

Protein Expr Purif. 2019 Nov:163:105451. doi: 10.1016/j.pep.2019.105451. Epub 2019 Jul 10.

Authors

Federica Palombarini¹, Francesca Ghirga², Alberto Boffi¹, Alberto Macone³, Alessandra Bonamore⁴

Affiliations

¹ Department of Biochemical Sciences, Sapienza University of Rome, P.le A. Moro 5, 00185, Rome, Italy.
² Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Viale Regina Elena 291, 00161, Rome, Italy.
³ Department of Biochemical Sciences, Sapienza University of Rome, P.le A. Moro 5, 00185, Rome, Italy. Electronic address: alberto.macone@uniroma1.it.
⁴ Department of Biochemical Sciences, Sapienza University of Rome, P.le A. Moro 5, 00185, Rome, Italy. Electronic address: alessandra.bonamore@uniroma1.it.

PMID: 31301427
DOI: 10.1016/j.pep.2019.105451

Abstract

Ferritin proteins are taking center stage as smart nanocarriers for drug delivery due to their hollow cage-like structures and their unique 24-meric assembly. Among all ferritins, the chimeric Archaeoglobus ferritin (HumFt) is able assemble/disassemble varying the ionic strength of the medium while recognizing human TfR1 receptor overexpressed in cancer cells. In this paper we present a highly efficient, large scale purification protocol mainly based on crossflow ultrafiltration, starting from fermented bacterial paste. This procedure allows one to obtain about 2 g of purified protein starting from 100 g of fermented bacterial paste. The current procedure can easily remove contaminant proteins as well as DNA molecules in the absence of expensive and time consuming chromatographic steps.

Keywords: Crossflow ultrafiltration; Diatomaceous earth; Ferritins; Humanized ferritin; Large scale purification.

MeSH terms

Archaeoglobus fulgidus / chemistry*
Archaeoglobus fulgidus / genetics
Cloning, Molecular
Escherichia coli / genetics
Ferritins / genetics
Ferritins / isolation & purification*
Humans
Recombinant Fusion Proteins / isolation & purification
Ultrafiltration / methods*

Substances

Recombinant Fusion Proteins
Ferritins