Page 6 - K. Development and regeneration

[K. Development and regeneration-4]

BAC based antibody expression platform using Tol2

transposon in CHO cells

Myeonguk Kuk¹, Joontae Park¹

¹Division of Life Sciences, Incheon National University, 연수구 22012, 대한민국

Engineering in the vector platform is commonly used to increase the production of recombinant protein in
mammalian cells. The current vector platform mainly uses a small size vector (1-20 kb), which enables to express
the protein easily and rapidly. However, this vector platform is very unstable because it is affected by the surrounding

environment of the locus, and the protein expression is greatly reduced as the generation passes. In this study, we

used a big size vector (100-500 kb), bacterial artificial chromosome (BAC), which enables to stably express the
protein over generation without being affected by the surrounding environment of the locus. First, we did bac
recombineering to incorporate the luciferase gene in the BAC encoding mouse Rosa26 locus. To facilitate the protein

expression, we also incorporated the Tol2 transposon system in Rosa26 BAC. We found that BAC was 8 μg, and
the ratio was optimal when BAC and transposase were 9:3. Furthermore, this platform was induced the more stable

expression of luciferase gene than the normal one. Taken together, this BAC with Tol2 platform may become a next
generation vector platform for the higher protein production in biopharmaceuticals.