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

  title={Engineered glycoforms of an antineuroblastoma IgG1 with optimized antibody-dependent cellular cytotoxic activity},
  author={Pablo Uma{\~n}a and Joel Jean-Mairet and Radmila Moudry and Hanspeter Amstutz and James Edwin Bailey},
  journal={Nature Biotechnology},
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… 

Non-fucosylated therapeutic antibodies as next-generation therapeutic antibodies

Non-fucosylated therapeutic antibodies exhibit the strongest and most saturable in vitro and ex vivo ADCC among such antibody variants with improved FcγRIIIa binding as those bearing naturally occurring oligosaccharide heterogeneities and artificial amino acid mutations, even in the presence of plasma IgG.

Antitumor Efficacy of Anti-GD2 IgG1 Is Enhanced by Fc Glyco-Engineering

A potent ADCC-enhanced antibody is described that, in comparison with Abs with altered affinities for Fc receptors, significantly improved the growth control of tumors expressing cancer-antigen GD2.

Function characterization of a glyco-engineered anti-EGFR monoclonal antibody cetuximab in vitro

The glyco-engineered EGFR mAb with more bisecting modifications and lower α-Gal content than the approved therapeutic antibody Erbitux shows improved functionality in vitro, and requires in vivo validations.

Modulation of IgG1 immunoeffector function by glycoengineering of the GDP‐fucose biosynthesis pathway

In an effort to significantly increase the potency of an anti‐CD20, IgG1 molecule, this effective glycoengineering strategy selectively targeted the de novo GDP‐fucose biosynthesis pathway of the host CHO cell line to generate >80% afucosylated IgG 1 resulting in enhanced FcγRIIIa binding and in vitro ADCC cell‐based activity.

Chemoenzymatic glycoengineering of intact IgG antibodies for gain of functions.

The glycoengineering approach described here provides a general platform to modulate the effector functions of IgG antibodies, enabling the optimization of therapeutic efficacy and gain of new functions of monoclonal antibodies and IVIG.

Production of afucosylated antibodies in CHO cells by coexpression of an anti‐FUT8 intrabody

It is demonstrated that expression of a membrane‐associated anti‐FUT8 intrabody engineered to reside in the endoplasmic reticulum and Golgi apparatus can efficiently reduce FUT8 activity and therefore the core‐fucosylation of the Fc N‐glycan of an antibody.

Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity

The production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila, is described and it is predicted that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans.

In Vivo Glycan Engineering via the Mannosidase I Inhibitor (Kifunensine) Improves Efficacy of Rituximab Manufactured in Nicotiana benthamiana Plants

The mannosidase I inhibitor kifunensine was applied to the Nicotiana benthamiana transient expression platform to produce an afucosylated anti-CD20 antibody and revealed a 14-fold increase in ADCC activity targeting the lymphoma cell line Wil2-S when compared with rituximab produced in the absence of kif unensine.

Engineered antibodies of IgG1/IgG3 mixed isotype with enhanced cytotoxic activities.

Findings suggest that the variant antibodies with IgG1/IgG3 chimeric constant regions and nonfucosylated oligosaccharides that possess dual-enhanced cytotoxic functions may be an improvement for the next generation of therapeutic antitumor antibodies.



Structural characterization of the N-glycans of a humanized anti-CD18 murine immunoglobulin G.

This study characterized the N-glycans of a humanized immunoglobulin G4 expressed in NS/O mouse myeloma cells and directed against the CD18 family of adhesion-promoting receptors on leukocytes, and indicated the presence of five oligomannoside-type structures and eight diantennary N-acetyllactosamine- type structures which are typical of mouse and human IgGs.

Glycosylation and biological activity of CAMPATH-1H expressed in different cell lines and grown under different culture conditions.

It is concluded that glycosylation of the antibody may be important in the clinical outcome of therapy.

Production and characterization of a mouse/human chimeric antibody directed against human neuroblastoma

A cell‐blnding inhibition assay showed that the specificity of the chimeric CE7 antibody (chCE7) is identical to that of the original CE7.

Depletion of B cells in vivo by a chimeric mouse human monoclonal antibody to CD20.

In vitro studies showed the ability of C2B8 to bind human C1q, mediate complement-dependent cell lysis of human B-lymphoid cell lines, and lyse human target cells through antibody-dependent cellular cytotoxicity and the possibility of using an "immunologically active" chimeric anti-CD20 antibody as an alternative approach in the treatment of B-cell lymphoma.

Variations in oligosaccharide-protein interactions in immunoglobulin G determine the site-specific glycosylation profiles and modulate the dynamic motion of the Fc oligosaccharides.

It is shown that two sets of IgG Fc glycoforms have quite different physical properties, interpreted to indicate a role for both the protein quaternary structure and specific protein-glycan interactions in determining the glycoform populations.

Anti-renal-cell carcinoma chimeric antibody G250 facilitates antibody-dependent cellular cytotoxicity with in vitro and in vivo interleukin-2-activated effectors.

  • J. SurfusJ. Hank P. Sondel
  • Biology, Medicine
    Journal of immunotherapy with emphasis on tumor immunology : official journal of the Society for Biological Therapy
  • 1996
In vitro data suggest that this chimeric antibody may improve efficacy of IL-2 therapy by targeting cytokine-activated effector cells directly to the tumor and facilitating in vivo ADCC.

Multiple interactions of IgG with its core oligosaccharide can modulate recognition by complement and human Fc gamma receptor I and influence the synthesis of its oligosaccharide chains.

It is suggested that noncovalent interactions of multiple amino acid residues of IgG with oligosaccharide residues that include the primary and secondary GlcNAc are necessary for optimal recognition of Igg by human Fc gammaRI and C1q.

Effect of glycosylation on antibody function: implications for genetic engineering.

Production of monoclonal antibodies in COS and CHO cells.