Expression of recombinant Alpha-1 antitrypsin in cho and cos-7 cell lines using lentiviral vector

ABSTRACT

In this study, in order to facilitate and accelerate the production of eukaryotic protein alpha 1-antitrypsin [AAT] with correct post-translational modifications, a protein production system based on the transduction of CHO and COS-7 cells using lentiviral vectors was developed. Human AAT cDNA was cloned into a replication- defective lentiviral vector. The transgene AATJred chimer was transferred to CHO and COS-7 cell lines using this vector and its expressions were visualized by fluorescent microscopy. The mRNA expression levels of the AAT genes were determined using Revearse Transcriptase-Polymerase Chain Reaction [RT-PCR] and its secretion into the medium by both cell types was determined using ELISA. The results show that by employing a lentiviral vector, efficient genetic loading of CHO and COS-7 cells with the AAT gene was achieved. In conclusion, by using a Lentivirus-based gene delivery system, large amounts of recombinant human AAT protein were expressed in both CHO and COS-7 cell lines. This expression system possesses key properties that ensure its application in the delivery of therapeutic genes into mammalian cultured cells