Effect of fortified amino acids on cell growth of a human hybrid cell line F2N78

초록

A human hybrid cell line named F2N78 was developed by somatic fusion between human embryonic kidney (HEK293) cells and Namalwa cells. HEK293 has been widely used as a human host for the effective transient expression of recombinant proteins. Namalwa cells are originated from Burkitt’s lymphoma infected by Epstein-Barr virus (EBV) and shows anti-apoptotic growth. As a result of cell fusion, F2N78 cell line inherited advantageous phenotypes such as ease to suspension, high transfection efficacy, EBV genome insertion in chromosome, and human-specific glycosylation. In order to be used in commercial processes, the formulation of the culture medium for F2N78 cell line has been investigated for optimal culture condition. In previous study, we studied media screening focused on the concentration of amino acids. To support cell growth, some amino acids were chosen as important factors. By comparing maximum viable cell density and specific growth rate, we screened significant factor from fortified amino acids, and finally isoleucine and cysteine were selected. The objective of this study is to find the best ratio between the selected amino acids which mitigate the negative effects of byproducts and improve cell growth. The concentration range of selected amino acids was set 1-fold, 2-fold, 3-fold and the experiments were designed by using 2-level fractional factorial design. With the addition of fortified level of isoleucine and cysteine, maximum viable cell density and specific growth rate could be enhanced, but viability was rapidly decreased. For further study, amino acid and vitamin screening focused on improving integral viable cell density (IVCD) will be investigated.