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Learn how tailored optimization strategies help overcome challenges in producing complex proteins using HEK-293 cells.
This poster highlights innovative solutions for enhancing protein yield and quality to advance research and therapeutic development.
Abstract
Obtaining high yields of pure and active difficult-to-produce recombinant proteins can be a real challenge and may require significant investment. The rapid screening of multiple production and purification conditions on a small scale is fundamental to identify the optimal strategy.
To select the best production conditions in HEK 293 EBNA cells, we first performed a small volume expression study to define optimal production time and ratio of DNA/transfection reagent. Moreover, we tested addition to the cell culture medium of various supplements (amino acids, inorganic salts, lipids…), aiming at boosting the expression level by varying medium composition. On identification of the best production conditions, several buffer compositions were screened at purification level to improve stability and solubility of the protein of interest. Different buffers, detergents, sugars, as well as α-helix stabilizers, and many more, were tested in a 96-well format for high-throughput.
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Authors
Isabelle Petit-Topin, Maxime Rochet, Isabelle Fixe, Flavien Carpentier, Alexandra Foucher, Nadia Normand
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Affiliation
Tebubio - 39 rue de Houdan - 78612 Le Perray-en-Yvelines - France
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