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Trans-splicing as a novel method to rapidly produce antibody fusion proteins |
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| Authors: | Ryohei Iwasaki Masaki Ihara Masayuki Kawakami |
| Affiliation: | a Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan b Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan c Lifescience Lab. R&D, Fujifilm Co., 577 Ushijima, Kaisei-machi, Ashigarakami-gun, Kanagawa 258-8577, Japan |
| Abstract: | To cultivate the use of trans-splicing as a novel means to rapidly express various antibody fusion proteins, we tried to express antibody-reporter enzyme fusions in a COS-1 co-transfection model. When a vector designed to induce trans-splicing with IgH pre-mRNA was co-transfected with a vector encoding the mouse IgM locus, the expression of VH-secreted human placental alkaline phosphatase (SEAP) as well as Fab-SEAP were successfully expressed both in mRNA and protein levels. Especially, the vectors encoding complementary sequence to Sμ as a binding domain was accurate and efficient, producing trans-spliced mRNA of up to 2% of cis-spliced one. Since Sμ sequence should exist in every IgH pre-mRNA, our finding will lead to the rapid production and analysis of various antibody-enzyme fusions suitable for enzyme-linked immunosorbent assay (ELISA) or antibody-dependent enzyme prodrug therapy (ADEPT). |
| Keywords: | TS, trans-splicing Fab, antibody fragment for antigen binding ELISA, enzyme-linked immunosorbent assay VH, variable fragment of H chain VL, variable fragment of L chain Fv, variable fragment scFv, single-chain Fv NP, 4-hydroxy-3-nitrophenacetyl |
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