Expression of GnTIII in a recombinant anti-CD20 CHO production cell line: Expression of antibodies with altered glycoforms leads to an increase in ADCC through higher affinity for FC gamma RIII
- PMID: 11410853
Expression of GnTIII in a recombinant anti-CD20 CHO production cell line: Expression of antibodies with altered glycoforms leads to an increase in ADCC through higher affinity for FC gamma RIII
Abstract
The gene encoding the rat glycosylation enzyme beta1-4-N-acetylglucosaminyltransferase III (GnTIII) was cloned and coexpressed in a recombinant production Chinese hamster ovary (CHO) cell line expressing a chimeric mouse/human anti-CD20 IgG1 antibody. The new cell lines expressed high levels of antibody and have growth kinetics similar to that of the parent. Relative QPCR showed the cell lines to express varying levels of mRNA. High-performance liquid chromatography (HPLC) analysis showed the enzyme to have added bisecting N-acetylglucosamine (GlcNAc) residues in most (48% to 71%) of the N-linked oligosaccharides isolated from antibody preparations purified from the cell lines. In an ADCC assay the new antibody preparations promoted killing of CD20-positive target cells at approximately 10- to 20-fold lower concentrations than the parent. This activity was blocked using an anti-Fc gamma RIII antibody, supporting the role of Fc gamma RIII binding in this increase. In addition, cell binding assays showed the modified antibody bound better to Fc gamma RIII-expressing cells. The increase in ADCC activity is therefore likely due to an increased affinity of the modified antibody for the Fc gamma RIII receptor.
Similar articles
-
Modulation of therapeutic antibody effector functions by glycosylation engineering: influence of Golgi enzyme localization domain and co-expression of heterologous beta1, 4-N-acetylglucosaminyltransferase III and Golgi alpha-mannosidase II.Biotechnol Bioeng. 2006 Apr 5;93(5):851-61. doi: 10.1002/bit.20777. Biotechnol Bioeng. 2006. PMID: 16435400
-
Establishment of FUT8 knockout Chinese hamster ovary cells: an ideal host cell line for producing completely defucosylated antibodies with enhanced antibody-dependent cellular cytotoxicity.Biotechnol Bioeng. 2004 Sep 5;87(5):614-22. doi: 10.1002/bit.20151. Biotechnol Bioeng. 2004. PMID: 15352059
-
Galactosylation of human IgG monoclonal anti-D produced by EBV-transformed B-lymphoblastoid cell lines is dependent on culture method and affects Fc receptor-mediated functional activity.Hum Antibodies Hybridomas. 1994;5(3-4):143-51. Hum Antibodies Hybridomas. 1994. PMID: 7756579
-
Human IgG Fc receptor heterogeneity: molecular aspects and clinical implications.Immunol Today. 1993 May;14(5):215-21. doi: 10.1016/0167-5699(93)90166-I. Immunol Today. 1993. PMID: 8517920 Review.
-
Engineering therapeutic antibodies for improved function.Biochem Soc Trans. 2002 Aug;30(4):487-90. doi: 10.1042/bst0300487. Biochem Soc Trans. 2002. PMID: 12196120 Review.
Cited by
-
Enhancing antibody-dependent cell-mediated cytotoxicity: a strategy for improving antibody-based immunotherapy.Antib Ther. 2018 Jun 24;1(1):7-12. doi: 10.1093/abt/tby002. eCollection 2018 Jun. Antib Ther. 2018. PMID: 33928217 Free PMC article. Review.
-
Enhancement of effector functions of anti-CD20 monoclonal antibody by increased afucosylation in CHO cell line through cell culture medium optimization.J Genet Eng Biotechnol. 2022 Oct 4;20(1):141. doi: 10.1186/s43141-022-00421-5. J Genet Eng Biotechnol. 2022. PMID: 36194313 Free PMC article.
-
Analytical characterization of ch14.18: a mouse-human chimeric disialoganglioside-specific therapeutic antibody.MAbs. 2012 Jan-Feb;4(1):84-100. doi: 10.4161/mabs.4.1.18566. MAbs. 2012. PMID: 22327432 Free PMC article.
-
Integrated glycomics and genetics analyses reveal a potential role for N-glycosylation of plasma proteins and IgGs, as well as the complement system, in the development of type 1 diabetes.Diabetologia. 2023 Jun;66(6):1071-1083. doi: 10.1007/s00125-023-05881-z. Epub 2023 Mar 13. Diabetologia. 2023. PMID: 36907892 Free PMC article.
-
Paracoccin from Paracoccidioides brasiliensis; purification through affinity with chitin and identification of N-acetyl-beta-D-glucosaminidase activity.Yeast. 2010 Feb;27(2):67-76. doi: 10.1002/yea.1731. Yeast. 2010. PMID: 19908201 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Other Literature Sources