Engineered glycoforms of an antineuroblastoma IgG1 with optimized antibody-dependent cellular cytotoxic activity

Umaña, Pablo; Jean‐Mairet, Joël; Moudry, R; Amstutz, Hanspeter; Bailey, James E.

doi:10.1038/6179

Cited by 722 publications

(520 citation statements)

References 25 publications

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“…Although it is difficult to assess the contribution of each of these mechanisms in their in vivo efficacy, clinical trial results support an important role of antibody-dependent cell-mediated cytotoxicity (ADCC) for both lymphomas and solid tumors (3). The affinity between the Fc portion of human IgG1 and FcgRIIIa (CD16a), an activating receptor mostly expressed by natural killer (NK) cells, monocytes, and macrophages, has a profound impact on ADCC exerted by antibodies (4,5). Correlation of clinical responses to mAb therapy with FcgRIIIa polymorphism has been observed with more favorable response in patients homozygous for the high-affinity FcgRIIIa (V158; refs.…”

Section: Introductionmentioning

confidence: 99%

Single-Domain Antibody–Based and Linker-Free Bispecific Antibodies Targeting FcγRIII Induce Potent Antitumor Activity without Recruiting Regulatory T Cells

Rozan

Cornillon

Pétiard

et al. 2013

Molecular Cancer Therapeutics

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Antibody-dependent cell-mediated cytotoxicity, one of the most prominent modes of action of antitumor antibodies, suffers from important limitations due to the need for optimal interactions with Fcg receptors. In this work, we report the design of a new bispecific antibody format, compact and linker-free, based on the use of llama single-domain antibodies that are capable of circumventing most of these limitations. This bispecific antibody format was created by fusing single-domain antibodies directed against the carcinoembryonic antigen and the activating FcgRIIIa receptor to human Ck and CH1 immunoglobulin G1 domains, acting as a natural dimerization motif. In vitro and in vivo characterization of these Fab-like bispecific molecules revealed favorable features for further development as a therapeutic molecule. They are easy to produce in Escherichia coli, very stable, and elicit potent lysis of tumor cells by human natural killer cells at picomolar concentrations. Unlike conventional antibodies, they do not engage inhibitory FcgRIIb receptor, do not compete with serum immunoglobulins G for receptor binding, and their cytotoxic activity is independent of Fc glycosylation and FcgRIIIa polymorphism. As opposed to anti-CD3 bispecific antitumor antibodies, they do not engage regulatory T cells as these latter cells do not express FcgRIII. Studies in nonobese diabetic/severe combined immunodeficient gamma mice xenografted with carcinoembryonic antigen-positive tumor cells showed that Fab-like bispecific molecules in the presence of human peripheral blood mononuclear cells significantly slow down tumor growth. This new compact, linker-free bispecific antibody format offers a promising approach for optimizing antibody-based therapies.

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Section: Introductionmentioning

confidence: 99%

Single-Domain Antibody–Based and Linker-Free Bispecific Antibodies Targeting FcγRIII Induce Potent Antitumor Activity without Recruiting Regulatory T Cells

Rozan

Cornillon

Pétiard

et al. 2013

Molecular Cancer Therapeutics

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“…Cell engineering of glycosylation patterns (Davies et al 2001;Natsume et al 2006;Shields et al 2002;Shinkawa et al 2003;Umañ a et al 1999;Warner 1999;Weikert et al 1999) and in vitro glycosylation (Butler 2005;Raju et al 2001) has provided new possibilities for biopharmaceutical design and optimization thus lowering immunogenicity effects, increasing stability and enhancing functional activity. Weikert et al (1999) engineered CHO cells that secreted either tissue necrosis factor receptor-IgG1 fusion protein (TNFR-IgG) or tissue plasminogen activator (TNK-tPA) to over-express a2,3-sialyltransferase and the resultant glycoproteins had a considerable longer pharmacokinetic mean residence time in rabbit models.…”

Section: Glycosylation Engineeringmentioning

confidence: 99%

“…Bisected oligosaccharides have been implicated in enhanced antibody dependant cellular cytotoxicity (ADCC) activity. When CHO cells were engineered to over express b1,4 N-acetyl glucosaminoyltransferase (GnTIII), the bisected oligosaccharide glycoforms, ADCC activity was increased 20-100 fold (Davies et al 2001;Umañ a et al 1999). A silencing approach has also been used in CHO cells to increase the antibody effector function of ADCC by introducing siRNA targeting a1,6 fucosyltransferase (FUT8) mRNA (Mori et al 2004).…”

Section: Glycosylation Engineeringmentioning

confidence: 99%

Using cell engineering and omic tools for the improvement of cell culture processes

et al. 2007

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Significant strides have been made in mammalian cell based biopharmaceutical process and cell line development over the past years. With several established mammalian host cell lines and expression systems, optimization of selection systems to reduce development times and improvement of glycosylation patterns are only some of the advances being made to improve cell culture processes. In this article, the advances pertaining to cell line development and cell engineering strategies are discussed. An overview of the cell engineering strategies to enhance cellular characteristics by genetic manipulation are illustrated, focusing on the use of genomics and proteomics tools and their application in such endeavors. Included in this review are some of the early studies using the 'omic' technique to understand cellular mechanisms of product synthesis and secretion, apoptosis, cell proliferation and the influence of the physicochemical environment. The article highlights the significance of integrating genomics and proteomics data with the vast amounts of bioprocess data for improved analysis of the biological pathways involved. Further improvements of the techniques and methodologies used are needed but ultimately, the new cell engineering strategies should provide great insight into the regulatory networks within the cell in a bioprocess environment and how to manipulate them to increase overall productivity.

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“…The contents of the dead and lysed cells are released to the cell culture supernatant, leading to contamination of the product. Most of the presently popular mammalian producer cell lines have been optimized with respect to technological requirements by adaptation and maintenance for extended periods of time under the appropriate culture conditions, while adaptation of the cellular physiology by genetic engineering is still in its infancy (Grabenhorst et al 1995;Monaco et al 1996;Irani et al 1999Irani et al , 2002Uman˜a et al 1999;Davies et al 2001;Fogolin et al 2004; for a review see Grabenhorst et al 1999). However, due to comparatively high costs and low productivities of cultured mammalian cells, there is a constant pressure to further improve the producer cell lines as well as the protein production processes.…”

Section: Introductionmentioning

confidence: 99%

Application of a Reversible Immortalization System for the Generation of Proliferation-Controlled Cell Lines

et al. 2004

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To employ physiological mechanisms to control cell growth primary cells were reversibly immortalized using the SV40 TAg. The cells showed a fibroblast-like morphology. When the expression of the TAg was turned off, the cells arrested in the G0/G1 cell cycle phase. The cell culture could be kept for over 1 week in the proliferation-controlled state while the growth arrest remained fully reversible. The regulation was highly efficacious in that the arrested cell population did not spontaneously resume growth, suggesting that in the absence of the immortalizing gene expression endogenous growth-control mechanisms can keep these cells in a viable state for a prolonged time. Recombinant protein expression increased in growth-controlled cells when compared to conventionally cultured cells. Analysis of a secreted pharmaceutical protein revealed high product integrity without any signs of degradation. Therefore, it is feasible to apply genetic regulation of cell immortalization to obtain proliferation-controlled cell lines and this technique may be of interest to generate novel biotechnological producer cells.Abbreviations: DMEM -Dulbecco's modified Eagle's medium; Dox -doxycycline; eGFP -enhanced green fluorescent protein; EPO -recombinant human erythropoietin; FCS -fetal calf serum; IRES -internal ribosomal binding site; LTR -long terminal repeat; MEF -murine embryonic fibroblast cell; neoNeomycin phosphotransferase; TAg -SV40 virus large T-antigen; w.t. -wild type

show abstract

Engineered glycoforms of an antineuroblastoma IgG1 with optimized antibody-dependent cellular cytotoxic activity

Cited by 722 publications

References 25 publications

Single-Domain Antibody–Based and Linker-Free Bispecific Antibodies Targeting FcγRIII Induce Potent Antitumor Activity without Recruiting Regulatory T Cells

Single-Domain Antibody–Based and Linker-Free Bispecific Antibodies Targeting FcγRIII Induce Potent Antitumor Activity without Recruiting Regulatory T Cells

Using cell engineering and omic tools for the improvement of cell culture processes

Application of a Reversible Immortalization System for the Generation of Proliferation-Controlled Cell Lines

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