TITLE : Production and Characterization of a Novel OX 40 Ligand for Clinical Use
نویسنده
چکیده
The cDNA or protein of interest, a Human-Fc TF20X40L fusion protein, was synthesized and integrated into a vector containing DHFR gene. This expression vector was then transfected into DG44, a DHFR deficient CHO cell line by electroporation. The transfected pool was selected with gradient MTX pressure before subcloning in semi-solid media. 1500 primary clones were screened for titer by ELISA. Top 10 primary clones were subcloned again with 20 subclones each picked and screened by ELISA. Fed-batch experiment was performed upon top 10 subclones in shake flask. Top 4 clones were chosen according to their titer both by ELISA and protein A HPLC confirmation. The fed-batch experiment of 4 top subclones was expanded to 1L bioreactors. Finally the 40 vials RCBs of top 4 subclones were established.
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