Purification of recombinant insect transferrin from large volumes of cell culture medium using high capacity Ni2+-dipicolylamine gel

Joy J. Winzerling, Daphne Q.D. Pham, Susan Kunz, Preminda Samaraweera, John H. Law, Jerker Porath

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

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 languageEnglish (US)
Pages (from-to)137-142
Number of pages6
JournalProtein Expression and Purification
Volume7
Issue number2
DOIs
StatePublished - Mar 1996

ASJC Scopus subject areas

  • Biotechnology

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