Trans-splicing as a novel method to rapidly produce antibody fusion proteins.
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).
Iwasaki
Ryohei
R
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Kiuchi
Hiroki
H
Ihara
Masaki
M
Mori
Toshihiro
T
Kawakami
Masayuki
M
Ueda
Hiroshi
H
D000469:Alkaline Phosphatase; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D019556:COS Cells; D002522:Chlorocebus aethiops; D004797:Enzyme-Linked Immunosorbent Assay; D058851:GPI-Linked Proteins; D005822:Genetic Vectors; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000235:genetics; D007075:Immunoglobulin M / Q000096:biosynthesis / Q000235:genetics; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D015202:Protein Engineering / Q000379:methods*; D012322:RNA Precursors / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D020040:Trans-Splicing
D000911:Antibodies, Monoclonal; D058851:GPI-Linked Proteins; D007140:Immunoglobulin Fab Fragments; D007075:Immunoglobulin M; D007527:Isoenzymes; D012322:RNA Precursors; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental