Expression of recombinant alpha-1 antitrypsin in CHO and COS-7 Cell lines using lentivirally vector

Abstract α1-antitrypsin (AAT) is a protease inhibitor that its primary function is to inhibit neutrophil elastase, a protease that can degrade all of the constituents of the pulmonary connective tissue matrix. AAT administration purified from blood is beneficial for AAT deficiency and cystic fibrosis and several other inflammatory diseases. A less expensive supply of AAT from recombinant producer can reduce patient treating cost compare to blood purified source. To facilitate and accelerate the production of eukaryotic proteins with correct post-translational modifications, we have developed a protein production system based on the transduction of CHO and COS-7 cells using lentivirally vectors. To accomplish this human AAT cDNA was cloned into a replication-defective lentiviral vector. The transgene AAT-Jred chimer was transferred to CHO and COS-7 cell line using this vector and its expression was visualized by fluorescent microscopy. The mRNA levels expression of AAT genes were determined using RT-PCR and its secretion from both cells into the medium was determined using ELISA.The results show that by employing a lentivirally vector efficient genetic loading of CHO and COS-7 cells with the AAT gene was achieved. The expression of the AAT gene in target cells was confirmed using RT-PCR. ELISA assay showed both CHO and COS-7 cells were active for secretion of AAT to medium.It has been concluded that by using a Lentivirus-based gene delivery system, we were able to express high amounts of recombinant human AAT protein in the CHO and COS-7 cell lines and this expression system possess key properties that ensure their employment to deliver therapeutic gene to mammalian cultured cells