Journal: Developmental and Comparative Immunology

Article Title: Structural and functional characterisation of ferret interleukin-2

doi: 10.1016/j.dci.2015.10.007

Figure Lengend Snippet: Size exclusion chromatography of purified ferret recombinant IL-2. Recombinant IL-2 was recovered by IMAC and fractionated on a Superdex 75 16/60 column. The column had previously been calibrated with BioRad gel filtration standards (chicken ovalbumin, 44 kDa; horse myoglobin, 17 kDa; and vitamin B12, 1.35 kDa). SDS-PAGE analysis (4–12% Bis-Tris NuPAGE) of peak fractions under both non-reduced and reduced conditions revealed: peak 1 contained high molecular weight host cell proteins; peak 2 – covalently -associated IL-2 dimer; and peak 3 – monomeric IL-2. Abbreviations: MW, molecular weight standards; SM, starting material; kDa, kilodalton; Vo, column void volume.

Article Snippet: Recombinant IL-2 was purified from 1.6 L of conditioned medium by immobilized metal ion affinity chromatography (IMAC; 5 mL HisTRAP HP column, GE Healthcare, Little Chalfont, UK) followed by gel filtration chromatography on a Superdex 75 16/60 column (GE Healthcare) equilibrated in phosphate-buffered saline (PBS) under conditions to minimise endotoxin contamination.

Techniques: Size-exclusion Chromatography, Purification, Recombinant, Filtration, SDS Page, High Molecular Weight, Molecular Weight