Abstract
We report the purification of secreted recombinant Manduca sexta transferrin from Spodoptera frugiperda (Sf9) cell culture medium in a single step using high capacity Ni2+-dipicolylamine (DPA)-Novarose gel. Although the original sample was highly diluted (~10 μg transferrin/ml medium) and the cell culture medium contained 10% surfactant (Pluronic F68) and a lipid emulsion, we were able to recover the recombinant transfertin (1 mg protein/100 ml) under gentle elution conditions with 70% yield at >90% homogeneity. This work demonstrates the versatility of immobilized metal ion affinity chromatography using a high metal ion capacity gel to purify a recombinant protein and illustrates the potential of this affinity technique for protein separations from large volumes of cell culture media that contain surfactants.
Original language | English (US) |
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Pages (from-to) | 137-142 |
Number of pages | 6 |
Journal | Protein Expression and Purification |
Volume | 7 |
Issue number | 2 |
DOIs | |
State | Published - Mar 1996 |
Externally published | Yes |
ASJC Scopus subject areas
- Biotechnology