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Lentiviral expression system for the purification of secreted proteins from human cell cultures | BMC Biotechnology | Full Text

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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. 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 293 T cells transfected with the corresponding plasmid. Secretion of sCD4 increased with the dose of the lentiviral vector up to a multiplicity of infection of 50. Exchanging the native signal peptide of sCD4 with the signal peptide of

Lentiviral expression system for the purification of secreted proteins from human cell cultures Methodology article Open access Published: 02 September 2016 Volume 16 , article number 66 ( 2016 ) Cite this article You have full access to this open access article Download PDF Save article View saved research BMC Biotechnology Aims and scope Submit manuscript Lentiviral expression system for the purification of secreted proteins from human cell cultures Download PDF Abstract Background Recombinant proteins of therapeutic use are ideally produced in human cells to ensure appropriate co- and post-

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