Establishment of FUT8 knockout Chinese hamster ovary cells: An ideal host cell line for producing completely defucosylated antibodies with enhanced antibody‐dependent cellular cytotoxicity
Top Cited Papers
- 17 August 2004
- journal article
- research article
- Published by Wiley in Biotechnology & Bioengineering
- Vol. 87 (5) , 614-622
- https://doi.org/10.1002/bit.20151
Abstract
To generate industrially applicable new host cell lines for antibody production with optimizing antibody‐dependent cellular cytotoxicity (ADCC) we disrupted both FUT8 alleles in a Chinese hamster ovary (CHO)/DG44 cell line by sequential homologous recombination. FUT8 encodes an α‐1,6‐fucosyltransferase that catalyzes the transfer of fucose from GDP‐fucose to N‐acetylglucosamine (GlcNAc) in an α‐1,6 linkage. FUT8−/− cell lines have morphology and growth kinetics similar to those of the parent, and produce completely defucosylated recombinant antibodies. FUT8−/−‐produced chimeric anti‐CD20 IgG1 shows the same level of antigen‐binding activity and complement‐dependent cytotoxicity (CDC) as the FUT8+/+‐produced, comparable antibody, Rituxan. In contrast, FUT8−/−‐produced anti‐CD20 IgG1 strongly binds to human Fcγ‐receptor IIIa (FcγRIIIa) and dramatically enhances ADCC to approximately 100‐fold that of Rituxan. Our results demonstrate that FUT8−/− cells are ideal host cell lines to stably produce completely defucosylated high‐ADCC antibodies with fixed quality and efficacy for therapeutic use.Keywords
This publication has 44 references indexed in Scilit:
- Structural Analysis of Human IgG-Fc Glycoforms Reveals a Correlation Between Glycosylation and Structural IntegrityPublished by Elsevier ,2003
- Lack of Fucose on Human IgG1 N-Linked Oligosaccharide Improves Binding to Human FcγRIII and Antibody-dependent Cellular ToxicityJournal of Biological Chemistry, 2002
- The Structure of a Human Type III Fcγ Receptor in Complex with FcJournal of Biological Chemistry, 2001
- Occurence of GDP-?-fucose: β-N-acetylglucosamine (Fuc to Asn-linked GlcNAc) α1,6-fucosyltransferases in porcine, sheep, bovine, rabbit and chicken tissuesBiochimica et Biophysica Acta (BBA) - General Subjects, 2000
- The α1-6-fucosyltransferase gene and its biological significanceBiochimica et Biophysica Acta (BBA) - General Subjects, 1999
- Targeting vector configuration and method of gene transfer influence targeted correction of theAPRT gene in Chinese hamster ovary cellsSomatic Cell and Molecular Genetics, 1993
- Targeted Introduction of a Diphtheria Toxin-Resistant PointMutation into the Chromosomal EF-2 Locus by in vivo Homologous Recombination.Cell Structure and Function, 1991
- Effect of gamma rays at the dihydrofolate reductase locus: Deletions and inversionsSomatic Cell and Molecular Genetics, 1986
- The control of glycoprotein synthesis: N-acetylgluocosamine linkage to a mannose residue as a signal for the attachment of L-fucose to the asparagine-linked N-acetylglucosamine residue of glycopeptide from α1-acid glycoproteinBiochemical and Biophysical Research Communications, 1976