Lentiviral expression system for the purification of secreted proteins from human cell cultures.

ABSTRACT:

Background

Recombinant proteins of therapeutic use are ideally produced in human cells to ensure appropriate co- and post-translational modifications. However, purification of secreted proteins from the culture media is impeded by low expression from transfected cell lines and the presence of serum proteins. Here we describe a simple and cost-effective approach based on lentiviral vector-mediated gene delivery and expression of a secreted His-tagged protein from human embryonic kidney 293 T cells and direct affinity chromatography purification from the cell culture media.

Results

Using a protein-based HIV entry inhibitor, soluble CD4 (sCD4), we demonstrated that 293 T cells transduced with a lentiviral vector mediated over 10-fold higher secretion of sCD4 in comparison to 29

SUBMITTER: Falkenhagen A 

PROVIDER: S-EPMC5009704 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature