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This galley proof is being listed electronically before publishing the final manuscript (It's not final version).

 
Antibody-secreting macrophages generated using CpG-free plasmid eliminate tumor cells through antibody-dependent cellular phagocytosis.
Eun Bi Cha1,2 (Graduate student), Keun Koo Shin1 (Researcher), Jinho Seo1,* (Researcher), Doo-Byung Oh1,2 (Principal Investigator)
1Environmental Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB),
2Biosystems and Bioengineering, University of Science and Technology (UST)
Abstract
The non-viral delivery of the gene into macrophages, known as hard-to-transfect cells, is a challenge. In this study, the microporation of a CpG-free and small-sized plasmid (pCGfd-GFP) showed high transfection efficiency, sustainable transgene expression, and good cell viability in the transfections of Raw 264.7 and primary bone marrow-derived macrophages. The non-viral method using the pCGfd vector encoding anti-EGFR single chain Fv fused with Fc (scFv-Fc) generated the macrophages secreting anti-EGFR scFv-Fc. These macrophages effectively phagocytized tumor cells expressing EGFR through the antibody-dependent mechanism, which was proved by experiments using EGFR-knockout tumor cells. Finally, peri-tumoral injections of anti-EGFR scFv-Fc-secreting macrophages were shown to inhibit tumor growth in the xenograft mouse model.
Abstract, Accepted Manuscript(in press) [Submitted on February 5, 2020, Accepted on February 28, 2020]
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