Tumour antigen targeted monoclonal antibodies incorporating a novel multimerisation domain significantly enhance antibody dependent cellular cytotoxicity against colon cancer

Jain, Ajay; Poonia, Bhawna; So, Edward; Vyzasatya, Ravi; Burch, Erin; Olsen, Henrik S.; Mérigeon, Emmanuel Y.; Block, David S.; Zhang, Xiaoyu; Schulze, Dan H.; Hanna, Nader; Twadell, William; Yfantis, Harris G.; Chan, Siaw-Lin; Cai, Ling; Strome, Scott E.

doi:10.1016/j.ejca.2013.06.009

Cited by 7 publications

(9 citation statements)

References 28 publications

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“…SDS-PAGE analysis proved that the resultant protein, GB4542, exists as a homodimer and higher-order multimers. Importantly, the differences in appearance between gels shown in this study and those previously reported by us are consistent with technological variances in the SDS-PAGE systems employed in the two reports, rather than a qualitative changes in GB4542 (28). As anticipated, SPR analysis and FACS-based cellular binding assays revealed that GB4542 demonstrated greatly enhanced avidity to the canonical FcgRs (Fig.…”

Section: Discussionsupporting

confidence: 78%

“…1A) (33-36). We previously used this strategy to generate recombinant murine IgG Fc multimers and a recombinant anti-human EGFR sAb (25,28). SDS-PAGE analysis proved that the resultant protein, GB4542, exists as a homodimer and higher-order multimers.…”

Section: Discussionmentioning

confidence: 99%

“…The amino acid sequence for rituximab was obtained from the ImMunoGeneTics database (http://www.imgt.org), and DNA fragments were synthesized at Blue Heron Biotechnology (Bothell, WA). G001, recombinant human IgG1 Fc; GL-2045, recombinant human IgG1 multimer; GB2500, recombinant anti-Her-2 mAb; and GB2542, recombinant anti-Her-2 sAb were all prepared by Gliknik (Baltimore, MD) (25,28).…”

Section: Generation and Production Of The Mab And Sabmentioning

confidence: 99%

“…The anti-CD20 mAb and GB4542 sAb constructs were generated essentially as described (28). The amino acid sequence for rituximab was obtained from the ImMunoGeneTics database (http://www.imgt.org), and DNA fragments were synthesized at Blue Heron Biotechnology (Bothell, WA).…”

Section: Generation and Production Of The Mab And Sabmentioning

confidence: 99%

“…Indeed, our studies revealed that anti-EGFR stradobodies (sAb), characterized by an anti-EGFR Fab linked to a multimerized Fc domain, mediated enhanced complement-dependent cytotoxicity (CDC) and Ab-dependent cellular cytotoxicity (ADCC) of EGFR-expressing colon cancer cell lines in vitro (28). However, we postulated that, like the stradomers, in the absence of Ag, the ability of the multimerized Fc fragments of the stradobodies to engage FcgRs and complement might prove tolerogenic.…”

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confidence: 99%

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Anti-CD20 Antibody with Multimerized Fc Domains: A Novel Strategy To Deplete B Cells and Augment Treatment of Autoimmune Disease

Zhang

Olsen

Chen

et al. 2016

The Journal of Immunology

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We developed a fully recombinant anti-CD20 protein derived from cDNA encoding one Fab domain, two IgG1 Fc regions, the IgG2 hinge, and an isoleucine zipper. This protein, called GB4542, contained both the homodimer and higher-order multimers. Binding studies revealed that GB4542 preferentially bound CD20+ cells yet also recognized CD20−FcγR+ PBMC. In contrast, a control mAb containing the identical Fab region, GB4500, failed to bind CD20−FcγR+ PBMC. Consistent with these findings, interactions between GB4542 and the canonical FcγRs had substantially lower KD values than correlate interfaces between GB4500 and these receptors. At low concentrations, GB4542 showed enhanced Ab-dependent cellular cytotoxicity, Ab-dependent cellular phagocytosis, and complement-dependent cytotoxicity compared with GB4500. However, at higher concentrations, an Fc analog of GB4542 inhibited anti-CD20 mAb–mediated B cell clearance through direct blocking of both Fc–FcγR interactions and C1q deposition on target cells. Furthermore, the higher-order multimer fraction of GB4542 demonstrated greater binding avidity with the canonical FcγRs and was associated with inhibitory effects observed in Ab-dependent cellular phagocytosis and complement-dependent cytotoxicity assays. These data suggest that GB4542 might have utility in the treatment of autoimmune diseases by combining both mAb-mediated B cell depletion and multimerized Fc-mediated tolerogenic effects.

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Section: Discussionsupporting

confidence: 78%

Section: Discussionmentioning

confidence: 99%

Section: Generation and Production Of The Mab And Sabmentioning

confidence: 99%

Section: Generation and Production Of The Mab And Sabmentioning

confidence: 99%

mentioning

confidence: 99%

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