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pmid: 19409879
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 V(H)-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 Smu as a binding domain was accurate and efficient, producing trans-spliced mRNA of up to 2% of cis-spliced one. Since Smu 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).
Recombinant Fusion Proteins, Genetic Vectors, 610, Antibodies, Monoclonal, Enzyme-Linked Immunosorbent Assay, Alkaline Phosphatase, GPI-Linked Proteins, Protein Engineering, Trans-Splicing, Isoenzymes, Immunoglobulin Fab Fragments, Mice, Immunoglobulin M, COS Cells, Chlorocebus aethiops, RNA Precursors, Animals, Humans
Recombinant Fusion Proteins, Genetic Vectors, 610, Antibodies, Monoclonal, Enzyme-Linked Immunosorbent Assay, Alkaline Phosphatase, GPI-Linked Proteins, Protein Engineering, Trans-Splicing, Isoenzymes, Immunoglobulin Fab Fragments, Mice, Immunoglobulin M, COS Cells, Chlorocebus aethiops, RNA Precursors, Animals, Humans
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