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

Nature Biotechnology volume 17, pages 176–180 (1999)Cite this article

Abstract

The glycosylation pattern of chCE7, an antineuroblastoma chimeric IgG1, was engineered in Chinese hamster ovary cells with tetracycline–regulated expression of β(1,4)–N–acetylglucosaminyltransferase III (GnTIII), a glycosyltransferase catalyzing formation of bisected oligosaccharides that have been implicated in antibody–dependent cellular cytotoxicity (ADCC). Measurement of the ADCC activity of chCE7 produced at different tetracycline levels showed an optimal range of GnTIII expression for maximal chCE7 in vitro ADCC activity, and this activity correlated with the level of constant region–associated, bisected complex oligosaccharides determined by matrix–assisted laser desorption/ionization time–of–flight mass spectrometry. The new optimized variants of chCE7 exhibit substantial ADCC activity and, hence, may be useful for treatment of neuroblastoma. The strategy presented here should be applicable to optimize the ADCC activity of other therapeutic IgGs.

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Figure 1: Representation of typical human Fc–associated oligosaccharide structures.
Figure 2: Tetracycline–regulated expression of c–myc–tagged GnTIII in CHO cells.
Figure 3: MALDI/TOF–MS spectra of neutral oligosaccharide mixtures from chCE7 samples produced by CHO–tet–GnTIII–chCE7 cell cultures differing in the concentration of tetracycline added to the media and, therefore, expressed the GnTIII gene at different levels.
Figure 4: N–linked oligosaccharide biosynthetic pathway leading to bisected complex and bisected hybrid oligosaccharides via GnTIII–catalyzed reactions.
Figure 5: (A) ADCC activity of different chCE7 samples.

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Acknowledgements

This research was supported by the Swiss Priority Program in Biotechnology (SPP BioTech). We would like to thank Ilse Novak for comparing the binding of different chCE7 samples to neuroblastoma cells, Heidi Ernst for DNA sequencing, and Naoyuki Taniguchi for providing us with the rat GnTIII cDNA.

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Authors and Affiliations

  1. ZLB Central Laboratory, Blood Transfusion Service, Swiss Red Cross, Bern, CH–3000 22, Switzerland

    Radmila Moudry & Hanspeter Amstutz

  2. Institute of Biotechnology, ETH Zürich, Zürich, CH–8093, Switzerland

    Pablo Umaña, Joël Jean–Mairet & James E. Bailey

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  1. Pablo Umaña
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Correspondence to James E. Bailey.

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Umaña, P., Jean–Mairet, J., Moudry, R. et al. Engineered glycoforms of an antineuroblastoma IgG1 with optimized antibody-dependent cellular cytotoxic activity. Nat Biotechnol 17, 176–180 (1999). https://doi.org/10.1038/6179

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