Figure 5
From: A streamlined implementation of the glutamine synthetase-based protein expression system
Comparison of the effectiveness of the IRES, dual promoter and T2A versions of the FcERα-eGFP-GS-pOPINEE12G vector for expressing FcERα in sorted cell populations, versus the yields obtainable following clone selection. A. FcERα expression from stable pOPINEE12G transfected cells was variable between triplicate transfections, with the IRES version giving slightly, but not significantly higher FcERα yields. Sorting for the brightest 30% considerably improved yield compared to unsorted populations of the same cells. B. FcERα yield from clonal cell lines obtained by conventional limiting dilution cloning. Clones 1 and 2 were selected at 35 μM MSX; clones 3-6 at 40 μM. FcERα titre was determined by competition ELISA on supernatant samples collected from 175 cm2 flasks after three weeks. Average yields from the triplicate sorted cell populations are indicated by the dotted lines; error bars indicate standard errors for triplicate ELISA readings.