M. Koerselman (Enschede, NL)

University of Twente Developmental BioEngineering

Presenter Of 1 Presentation

Poster Osteoarthritis

P144 - Controlled Release of Heavy Chain Only Antibodies Targeting IL-1R to Block IL-1 Signalling

Presentation Topic
Osteoarthritis
Date
13.04.2022
Lecture Time
09:30 - 09:30
Room
Exhibition Foyer
Session Name
7.3 - Poster Viewing / Coffee Break / Exhibition
Session Type
Poster Session
Disclosure
No Significant Commercial Relationship

Abstract

Purpose

Intra-articularly injected biologicals are limited in disease modification by their short in vivo half-life, poor stability and loss of bioactivity. Anakinra, which is a modified recombinant version of IL-1RA is such a therapeutic protein suffering from these limitations. Here, we report a potential alternative for Anakinra, consisting of the variable domain of heavy chain only antibodies (VHHs or nanobodies). The aim is to have a long-term controlled release of bioactive VHH to block IL-1 signaling in the intra-articular space.

Methods and Materials

Several clones of VHHs targeting IL-1R were obtained after immunization of Llama’s, phage library construction and panning rounds. ELISAs and Surface Plasmon Resonance imaging (SPRi) was used to characterize the binding affinity of the VHHs. Bioactivity of the VHHs was determined using a HEK293t cell line stably transfected with an NFκB inducible reporter construct after co-stimulation with IL-1β. As a positive control, a biosimilar of IL-1RA was used. In vitro release studies were carried out using slow release systems composed of various compositions of PCLA-PEG-PCLA triblock co-polymers.

Results

We obtained clones of VHHs targeting IL-1R that had an even higher binding affinity than IL-RA and were more potent than IL-RA in neutralizing Il-1B (fig. 1 and 2). As a next step, these two VHHs were incorporated in slow release systems. Over a time span of 60 days, samples were taken and we observed a sustained release in two compositions. Simultaneously, biological activity was confirmed over time.

icrs2_1.pngicrs.png

Conclusion

We obtained two high affinity IL-1R binding VHHs that could block IL-1 signaling. It is possible to formulate these VHHs in biomaterials for prolonged release without losing biological activity. These VHH formulations are a cost-effective alternative for intra-articular IL-1RA injections.

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Presenter Of 1 Presentation

Osteoarthritis

P144 - Controlled Release of Heavy Chain Only Antibodies Targeting IL-1R to Block IL-1 Signalling

Abstract

Purpose

Intra-articularly injected biologicals are limited in disease modification by their short in vivo half-life, poor stability and loss of bioactivity. Anakinra, which is a modified recombinant version of IL-1RA is such a therapeutic protein suffering from these limitations. Here, we report a potential alternative for Anakinra, consisting of the variable domain of heavy chain only antibodies (VHHs or nanobodies). The aim is to have a long-term controlled release of bioactive VHH to block IL-1 signaling in the intra-articular space.

Methods and Materials

Several clones of VHHs targeting IL-1R were obtained after immunization of Llama’s, phage library construction and panning rounds. ELISAs and Surface Plasmon Resonance imaging (SPRi) was used to characterize the binding affinity of the VHHs. Bioactivity of the VHHs was determined using a HEK293t cell line stably transfected with an NFκB inducible reporter construct after co-stimulation with IL-1β. As a positive control, a biosimilar of IL-1RA was used. In vitro release studies were carried out using slow release systems composed of various compositions of PCLA-PEG-PCLA triblock co-polymers.

Results

We obtained clones of VHHs targeting IL-1R that had an even higher binding affinity than IL-RA and were more potent than IL-RA in neutralizing Il-1B (fig. 1 and 2). As a next step, these two VHHs were incorporated in slow release systems. Over a time span of 60 days, samples were taken and we observed a sustained release in two compositions. Simultaneously, biological activity was confirmed over time.

icrs2_1.pngicrs.png

Conclusion

We obtained two high affinity IL-1R binding VHHs that could block IL-1 signaling. It is possible to formulate these VHHs in biomaterials for prolonged release without losing biological activity. These VHH formulations are a cost-effective alternative for intra-articular IL-1RA injections.

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