Antitumor activity of PEGylated Fab-fragments of GD2-specific antibody
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1
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Russia
Monoclonal antibodies against tumor-associated ganglioside GD2 are attractive tools for researchers and clinicians as an agent for anti-cancer immunotherapy. Despite some success of their application, there are a number of side effects associated with Fc-fragment of GD2-mAb. Recently, it was shown that the GD2-mAb has direct strong cytotoxic activity against tumor cells expressing GD2. The fact that these effects did not associated with antibody’s Fc-fragment allows the use of active antibody fragments without Fc-region (Fab-fragments). Purpose of the study is to evaluate the possibility of applying Fab-fragments of GD2-mAb for tumor therapy.
In our work, we have obtained purified fraction of Fab-fragments from GD2-mAbs. Using flow cytometry assay, we showed their antitumor effects on GD2-positive tumor cell lines. The main drawback of smaller Fab-fragments in comparison to whole antibodies is their drastically shorter elimination half-life. In order to increase the half-life of Fab-fragment GD2-mAb we produced their site-specific PEGylation. Maleimide-based reaction allows to covalently attach PEG (from 1 to 3 molecules per Fab) to cysteines of Fab-fragment far away from the epitope binding region. Using ELISA, flow cytometry assays and confocal microscopy, we have shown that PEGylation does not lead to significant changes in the binding of GD2 compared to unmodified Fab-fragments. Thus the use of this method has allowed us to maintain the activity of Fab-fragment GD2-mAb.
We have established a syngeneic mouse model of GD2-positive tumor. Using different criteria for evaluating of tumor progression, we analyzed antitumor effects of full-size GD2-mAb, their Fab-fragments and PEGylated Fab-fragments of GD2-mAb.
Acknowledgements
This research was supported by Federal program "Scientific and scientific-pedagogical personnel of innovative Russia" (2009-2013), state contract number 8165.
Keywords:
Gangliosides,
GD2,
Immunotherapy,
site-specific pegilation,
Fab-fragments
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Immune receptors and signaling
Citation:
Doronin
II,
Kholodenko
IV,
Molotkovskaya
IM and
Kholodenko
RV
(2013). Antitumor activity of PEGylated Fab-fragments of GD2-specific antibody.
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.00193
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Received:
11 Mar 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Mr. Igor I Doronin, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, Moscow, 117997, Russia, doronin@lactocore.com
Mr. Roman V Kholodenko, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, Moscow, 117997, Russia, khol@mail.ru