Affinity-controlled capture and release of engineered monoclonal antibodies by macroporous dextran hydrogels using coiled-coil interactions

ABSTRACT

Long-term delivery is a successful strategy used to reduce the adverse effects of monoclonal antibody (mAb)-based treatments. Macroporous hydrogels and affinity-based strategies have shown promising results in sustained and localized delivery of the mAbs. Among the potential tools for affinity-based delivery systems, the de novo designed Ecoil and Kcoil peptides are engineered to form a high-affinity, heterodimeric coiled-coil complex under physiological conditions. In this study, we created a set of trastuzumab molecules tagged with various Ecoil peptides and evaluated their manufacturability and characteristics. Our data show that addition of an Ecoil tag at the C-termini of the antibody chains (light chains, heavy chains, or both) does not hinder the production of chimeric trastuzumab in CHO cells or affect antibody binding to its antigen. We also evaluated the influence of the number, length, and position of the Ecoil tags on the capture and release of Ecoil-tagged trastuzumab from macroporous dextran hydrogels functionalized with Kcoil peptide (the Ecoil peptide-binding partner). Notably, our data show that antibodies are released from the macroporous hydrogels in a biphasic manner; the first phase corresponding to the rapid release of residual, unbound trastuzumab from the macropores, followed by the affinity-controlled, slow-rate release of antibodies from the Kcoil-functionalized macropore surface.

KEYWORDS:

• Affinity
• Coiled-coils
• Hydrogels
• Monoclonal antibodies
• Sustained release

Acknowledgments

This work was supported by the Natural Sciences and Engineering Research Council of Canada via the NSERC-CREATE PrEEmiuM program (S.F.B., R.O.) and the NSERC Discovery program (N.V.) and by the Fonds de Recherche du Québec – Nature et Technologies (R.O., X.B.). E.P. is grateful for the funding received from the European Research Council (Grant Agreement No. 851179), the Consellería de Educación, Universidade e Formación Profesional, Xunta de Galicia (ED431C 2018/39, ED431C 2022/39 and 508/2020), and the Agencia Estatal de Investigación for her Ramón y Cajal contract (RYC2019-027199-I).

The authors also acknowledge financial support from the TransMedTech Institute and its main funding partner, the Canada First research and excellence fund (R.O.), from Xunta de Galicia, Axudas de Apoio á Etapa de Formación Predoutoral, ED481A-2021/008 (I.O.G.), and from the Canada Research Chair funding program (X.B.).

The authors would also like to thank Anik Chevrier and Pr. Marc Lavertu (Chemical Engineering, Polytechnique Montréal — confocal microscopy), and Catherine Forest-Nault and Jimmy Gaudreault (Chemical Engineering, Polytechnique Montréal — SPR measurements) for their experimental assistance. This is NRC publication #NRC-HHT_53745

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/19420862.2023.2218951

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

The work was supported by the Agencia Estatal de Investigación [RYC2019-027199-I]; Canada First Research Excellence Fund [TransMedTech]; Canada Research Chairs European Research Council [851179]; Fonds de recherche du Québec – Nature et technologies Natural Sciences and Engineering Research Council of Canada [NSERC-CREATE PrEEmiuM program]; Xunta de Galicia [ED431C 2018/39, ED431C 2022/39, 508/2020]; Xunta de Galicia [ED481A-2021/008]