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Engineering Chinese hamster ovary cells to maximize effector function of produced antibodies using FUT8 siRNA
Author(s) -
Mori Katsuhiro,
KuniKamochi Reiko,
YamaneOhnuki Naoko,
Wakitani Masako,
Yamano Kazuya,
Imai Harue,
Kanda Yutaka,
Niwa Rinpei,
Iida Shigeru,
Uchida Kazuhisa,
Shitara Kenya,
Satoh Mitsuo
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20326
Subject(s) - chinese hamster ovary cell , antibody dependent cell mediated cytotoxicity , antibody , small interfering rna , microbiology and biotechnology , fucosyltransferase , transfection , recombinant dna , biology , agglutinin , cell culture , chemistry , biochemistry , gene , monoclonal antibody , immunology , lectin , genetics
We explored the possibility of converting established antibody‐producing cells to cells producing high antibody‐dependent cellular cytotoxicity (ADCC) antibodies. The conversion was made by constitutive expression of small interfering RNA (siRNA) against α1,6 fucosyltransferase (FUT8). We found two effective siRNAs, which reduce FUT8 mRNA expression to 20% when introduced into Chinese hamster ovary (CHO)/DG44 cells. Selection for Lens culinaris agglutinin (LCA)‐resistant clones after introduction of the FUT8 siRNA expression plasmids yields clones producing highly defucosylated (≈60%) antibody with over 100‐fold higher ADCC compared to antibody produced by the parental cells (≈10% defucosylated). Moreover, the selected clones remain stable, producing defucosylated antibody even in serum‐free fed‐batch culture. Our results demonstrate that constitutive FUT8 siRNA expression can control the oligosaccharide structure of recombinant antibody produced by CHO cells to yield antibodies with dramatically enhanced ADCC. © 2004 Wiley Periodicals, Inc.