title: Production and purification of recombinant human hepcidin-25 with authentic N and C-termini.
abstract: Hepcidin was first identified as an antimicrobial peptide present in human serum and urine. It was later demonstrated that hepcidin is the long-sought hormone that regulates iron homeostasis in mammals. Recombinant human Hepcidin-25 (Hepc25) was expressed in Pichia pastoris using a modified version of the pPICZαA vector. Hepc25 was then purified by a simple two-step chromatographic process to obtain 1.9 mg of soluble recombinant human Hepc25 per liter of culture at 96% purity. The sequence of Hepc25 and the presence of four disulfide bridges were confirmed by mass spectrometry analyses, and the recombinant Hepc25 exhibited antibacterial activity. This protocol of production and purification is the first step toward the production of human Hepc25 at a greater scale.
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Vignesh Narasimhan
authors.initials: VN
authors.lastname: Janakiraman
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Cabanne
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Wilfrid
authors.initials: W
authors.lastname: Dieryck
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Hocquellet
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Joucla
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Le Senechal
authors.affiliation: Univ Bordeaux, CNRS, CBMN, UMR5248, F-33000 Bordeaux, France.
authors.forename: Stephane
authors.initials: S
authors.lastname: Chaignepain
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Patricia
authors.initials: P
authors.lastname: Costaglioli
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: Santarelli
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France. Electronic address: bertrand.garbay@ipb.fr.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Garbay
authors.affiliation: Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Abdelmajid
authors.initials: A
authors.lastname: Noubhani
mesh_terms: D002846:Chromatography, Affinity; D064451:Hepcidins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D064451:Hepcidins; D011994:Recombinant Proteins; C545475:hepcidin 25, human
keywords: Expanded bed adsorption; Hepcidin; IMAC; P. pastoris; Recombinant expression