The Steyaert lab pioneered the use of nanobodies in X-ray crystallography, aiming at the highest-hanging fruits of structural biology including membrane proteins, amyloidogenic proteins, and now also (transient) multiprotein complexes. Recent work focuses on exploiting nanobodies for drug discovery on difficult targets like GPCRs and on the engineering of nanobodies to be used in single-particle cryo-EM and OMICs applications.

Tectonic Therapeutic is a preclinical stage biotechnology company transforming the discovery of novel GPCR-targeted therapies. Our GEODe platform is a suite of discovery tools comprising biochemistry techniques, protocols that enable presentation of GPCR antigens in the correct conformation to enable successful discovery campaigns using yeast-display antibody libraries. Our antigen engineering approaches boost receptor, bias receptor conformation to facilitate agonist/antagonist selections and improve receptor thermostability for selection condition compatibility.

Membrane proteins are still the greatest challenge when it comes to antibody generation against these complex targets. Whilst there are numerous options for immunisation and screening, there is no one method that is guaranteed for every target and thus multiple strategies have to be used. Here I present some examples of antibody discovery against challenging targets (GPCRs , Ion channels) done at Glaxosmithkline.

Even well-characterized cell surface receptors can pose challenges in selective targeting and efficient disease modulation and at the same time offer opportunities for innovation. In this talk, I will present antibody discovery at Novo Nordisk and give a case story of how we have generated antibodies targeting a receptor-ligand complex for the treatment of type 2 diabetes.

ConfoBodies are camelid-derived immunoglobulin single variable domains that stabilize disease-relevant pharmacological states of GPCRs. Such tools are used for in vitro screening immediately triaging for compounds with the pharmacology of interest and to enable structure-guided drug discovery. Confo Therapeutics has established large naive and synthetic VHH repertoires. Without the need for affinity maturation, active state stabilizing ConfoBodies from such non-immune repertoires have been identified that enable agonist compound screening and structural biology.

Multi-pass transmembrane proteins represent some of the most important drug target classes across a wide range of therapeutic areas. An update on antibody-based therapeutics in the GPCR, ion channel, and transporter pipeline will be provided outlining the breadth and diversity of the target landscape, progress in preclinical and clinical development, including next-generation modalities.

GPCRs and Ion Channels are important drug targets yet are notoriously difficult to work with. We’ll present the direct extraction of GPCRs from crude cells as well as case studies on our latest developments, including CXCR4 oligomers that can be reconstituted into Salipro-CXCR4 nanoparticles, enabling new possibilities for development of therapeutic antibodies, including phage display, immunisation, B-cell sorting and antibody characterisation with SPR and high-resolution cryoEM.

Resolution of inflammation is elicited by proresolving lipids activating GPCRs. ChemR23 is overexpressed in inflamed colon tissues of severe IBD patients unresponsive to anti-TNF or integrins therapies and associated with mucosal neutrophil accumulation. We generated an anti-ChemR23 mAb for Resolvin E1-like activity capable to repolarize human Macrophages and limits Neutrophil-mediated inflammation and NETosis. This mAb triggered resolution in ongoing chronic colitis models with significant decrease in tissue fibrosis. 

To generate an effective antibody-based biotherapeutic it is important to first produce a sample of the relevant antigen in a form that mimics the disease state. In the most basic form this means generating antigens that are folded, however in more complex systems may also mean antigens that are based on protein complexes. The challenge of native antigen generation becomes even more complex for membrane proteins which are structurally labile. Over the last 10 years we have demonstrated that the Styrene Maleic Acid Copolymer Lipid Particle (SMALP), provides a simple method for generating such membrane protein antigens. I will show how SMALPs can be used to generate multiprotein antigens in their native lipid environment with native conformations and activities. This makes them ideally suited to biotherapeutic antibody generation.