DEVELOPMENT OF NEW HEK-293 CELL LINES FOR THERAPEUTIC PROTEIN PRODUCTION

EDUARDO MUFARREGE; EVANGELINA BENIZIO; CELESTE RODRIGUEZ; MARINA ECHEVERRIGARAY; RICARDO KRATJE; CLAUDIO PRIETO

Buenos Aires

Congreso; SAIB - Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2013

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Recombinant protein production in animal cells shows numerous advantages, such as folding and post-translational modifications (phosphorylation and glycosylation). However, the main limitation resides in the difficulty to produce them in high levels. To solve this drawback, in the present study, we aimed to generate HEK- 293 cell lines over-expressing a gene expression key protein designated here as ?P? (for confidential purposes). This protein is involved in different mRNA metabolism steps, such as stability, transport and translation. These new cell lines were tested for recombinant gene expression. Firstly, EGFP (enhanced green fluorescence protein) expression analysis revealed that the HEK-P cell line produced a 2-fold increase in EGFP levels compared with wild type cells. Then, by means of a cell cloning strategy, individual clones from the HEK-P cell line that showed a 4-fold increase in the fluorescence levels were identified. Interestingly, these clones have shown similar behavior when two therapeutic protein productions (rhEPO and rhIFN-á) were evaluated. Additionally, other parameters were considered, including growth characteristics, morphology, glucose consumption and lactate and ammonium production. Taken together, these results represent a significant advance in the development of optimized cell lines widely used in the pharmaceutical industry.