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

BMC Biotechnol.. 2016; 
FalkenhagenAlexander,AsadSabah,ReadStanley E,JoshiSa
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DNA Sequencing … The gene encoding sCD4 (amino acids 1-209 of CD4, UniProtKB P01730, followed by GGGSGAGCCPGCC HHHHHH) and the different signal peptides were synthesized and sequenced by Genscript (Piscataway, United States) … Get A Quote

摘要

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.

关键词

His-tag,Human cells,Lentiviral vector,Purification,Secreted proteins,Soluble