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Fernanda Santos Arthuso

Fernanda Santos Arthuso

Institute of Nuclear and Energy Research – IPEN/CNEN
University of São Paulo
Brazil

Title: Adaptation of human prolactin secreting CHO cells to growth in suspension

Biography

Fernanda Santos Arthuso obtained the Biology degree from the University Paulista, has completed her master degree in science at University of São Paulo and is a PhD student at the Biotechnology Center of the Institute for Nuclear and Energy Research. She has experience in Biotechnology concerning recombinant protein expression in bacteria and CHO cells. She acts primarily in the following fields: synthesis, purification and characterization of recombinant pituitary hormones.

Abstract

Production of recombinant protein in suspension adapted cells show a great industrial-pharmaceutical interest, especially because of reduction of cultivation handling and improvement in scale enlargement as well as of increasing volumetric productivity. We initially developed the expression of recombinant human prolactin (hPRL) by Chinese Hamster Ovary cells (CHO) in monolayer culture dish depending on fetal bovine serum (FBS) in the medium for an efficient growth. In this work we described the development of a fast and practical protocol for adapting these CHO cells to suspension growth, in ~40 days, in serum-free medium. High cell densities, up to 4 x 106 cells per mL, were obtained in spinner flask cultures and a stable and continuous production process was observed for at least 30 days. Two harvesting strategies were set up, 50 or 100% of the total conditioned medium being collected daily and replaced by fresh culture medium. The volumetric productivity was 5–7 μg hPRL/ml, as determined directly in the collected medium via reversed-phase HPLC (RP-HPLC). We also carried out laboratory scale production and the corresponding purification and characterization of this protein.