Figure 1

Tg10 mAb production in mice subjected to intramuscular electrotransfer. (A) Tg10 mAb-expressing vectors used for electroporation. pCOR-κTg10 and pCOR-hTg10, pCOR-derived vectors expressing the Tg10 light (κTg10) and Tg10 heavy (hTg10) chains cDNAs under the control of the CMV promoter, respectively. CMV-tTA and CMV-rtTA express tTA and rtTA transactivators under the transcriptional control of the CMV promoter, respectively. In tetO-Tg10, κTg10 and hTg10 are expressed from a monocistronic expression cassette owing to the poliovirus internal ribosome entry sequence (IRES) placed under the cis-control of a minimal CMV promoter linked to multiple copies of the bacterial tetO operator [24]. (B): in vivo Tg10 production after electroporation of pCOR-derived vectors. Ten 4 week-old C57Bl6 mice were divided into 2 groups and injected intramuscularly in the tibialis anterior with either 20 μg of pCOR-κTg10 plus 20 μg of pCOR-hTg10 (mice 1 to 5) or a saline solution taken as a negative control. Electroporation was then performed as described in [15]. Blood samples were withdrawn at the indicated time points post-electroporation and serum Tg10 levels were assayed by ELISA [4]. (C): Regulatable in vivo Tg10 mAb production. 4 week-old C3H mice were divided into 3 groups of 5 animals and injected intramuscularly in the tibialis anterior with either a saline solution (not shown), 100 μg of tetO-Tg10 (not shown) or 100 μg of CMV-tTA plus 100 μg of tetO-Tg10 (mice 27 to 31). Electroporation was then performed as described in [15]. Doxycycline was added or removed from mice drinking water at indicated times. Tg10 levels in serum samples taken at different time points post-electroporation were assayed by ELISA.