Engineering and selecting an antibody for high affinity and potency can sometimes come at the expense of a desirable developability profile. In this presentation, we will describe complementary methods to improve affinity and developability whilst maintaining function in a panel of C7 antibodies. We identified antibodies that had desired human, cynomolgus monkey, and/or rat cross-reactive profiles and each had a distinct, novel mechanism of C7 inhibition. The lead antibody was effective in preventing experimental Myasthenia Gravis in rats in both prophylactic and therapeutic dosing regimens, and this provides therapeutic options for treatment of MG patients.

Proteases play an essential role in maintaining tissue homeostasis and aberrant activation leads to cancer or inflammatory diseases. The development of selective and potent protease inhibitory antibodies offers tremendous therapeutic potential. I will outline two case studies where we develop inhibitory antibodies with picomolar affinities and new modes of action. We leverage these antibodies to reveal the role of proteases as primary drivers of inflammatory disease in several mouse models.

Aberrant signaling of the tumor necrosis factor receptor family has significant detrimental effects in multiple disease states. Ligand competition impacts multiple pathways, causing an array of side effects, and is challenged by native potency and high local concentrations. Synthetic scaffolds were engineered to bind receptors (separately TNFR1 and DR5) and inhibit signaling and downstream processes without competing for native ligand binding. Mechanistic impacts of receptor conformation were evaluated.

• How can we address the challenges of difficult targets?
• Can we still improve the next generation of repertoires?
• What are the Impacts of machine learning and bioinformatics?
• What is the niche for academic vs. industry research?

• When to develop biologics instead of small molecules?
• Can AI help identify the best targets for biologics or small molecules?
  
• What are the commercial considerations for the development of biologics vs small molecules?
  

ARGX-119 is an agonistic anti- muscle-specific kinase (MuSK) antibody, derived from the SIMPLE antibody platform, with broad potential in neuromuscular diseases. Congenital myasthenia (CM) is a devastating neuromuscular disease and mutations in DOK7 are a major cause of CM. We developed agonist antibodies against MUSK and show that these antibodies restored neuromuscular synapse formation and prevented neonatal lethality and late-onset disease in mouse model for DOK7 CM.

There is a significant need for targeting new pathological proteins to develop novel therapies for neurodegenerative diseases. Metalloproteinases (MPs) play key physiological and pathological roles in the central nervous system by regulating signaling pathways during neuroinflammation, blood-brain barrier disruption, or synaptic dysfunction which makes them great targets for developing therapeutics for neurodegenerative diseases. Due to multifaceted role of MPs in cellular function, it is desired to use protein engineering and design approaches to generate MP inhibitors that inhibit specific MPs upregulated in the brain tissue and other related tissues of AD patients.

Proteases are important drug targets but finding their specific mAbs not only binding but also inhibiting is often a bottleneck. We develop streamlined methodologies coupling novel synthetic library designs with functional selections, and generated panels of potent and specific mAbs inhibiting numerous proteases of biomedical importance. Exhibiting efficacy in mouse models of cancers, neuropathic pains, obesity, and stroke, our mAbs show therapeutic promises as safe and effective protease inhibitors.

The Notch pathway is conserved in all metazoans, but safely drugging this target has remained an elusive challenge. Herein we describe the discovery and characterization of a Jag-ligand selective anti-Notch2 antibody that binds a unique epitope on Notch2. We demonstrate this selectivity via systemic administration of this antibody, which causes selective transdifferentiation of the mouse airway without causing DLL-dependent systemic phenotypes.

Antibodies specific to oxidative post-translationally modified insulin (oxPTM-INS) are present in most individuals with type 1 diabetes (T1D), even before the clinical onset. We investigated the antibody response to oxPTM-INS neoepitope peptides (oxPTM-INSPs) and evaluated their ability to stimulate humoral and T cell responses in T1D. We combined size-exclusion chromatography, LC-MS/MS, ELISA, and T cells proliferation assays to identify the oxPTM-INSPs that are involved in the immunopathogenesis of T1D.

We have optimized three separate antibodies for increased potency and other beneficial characteristics. A poorly-expressed p95-binding mouse mAb was optimized for nearly an order of magnitude improvement in potency and nearly two orders of magnitude increased expression. A humanized tyrosine kinase receptor (TKR) agonistic antibody with modest cross-reactivity to the murine TKR was improved in potency by nearly two orders of magnitude against both species. Finally, a mouse mAb against an undisclosed soluble extracellular protein was humanized and optimized for potency using a designed yeast display library and by approximately two orders of magnitude but maintained the relevant selectivity.

We have been advancing “muco-trapping” mAbs, which crosslink pathogens to mucins via polyvalent Fc-mucin bonds, as inhaled therapy of acute respiratory infections (ARIs). We will present data on preclinical experience in small and large animal models, execution of IND-enabling studies demonstrating feasibility of stably nebulizing mAbs, and our clinical safety and PK findings. Our progress underscores the promise of inhaled muco-trapping mAb platform for early treatment of various ARIs.

PCSK9 and ANGPTL3 are two targets that have FDA-approved monoclonal antibodies against them, both causing significant reductions in LDL cholesterol and triglycerides. RNA therapies, including small interfering RNAs (siRNAs) and antisense oligonucleotides (ASOs), work intracellularly to inhibit protein translation. This prevents key proteins of interest, such as PCSK9, ANGPTL3, APOC3, or Lp(a), from being produced in the cell, reducing serum levels by as much as 95%.

AMG 133 is a novel gastric inhibitory polypeptide receptor (GIPR) antagonistic antibody and GLP-1 peptide conjugate that exhibited remarkable body weight loss and significant improvement of metabolic parameters in preclinical models. In human Phase 1 trial AMG 133 demonstrated weight loss and tolerability. The design, conjugation process, and preclinical activity will be discussed.

A single administration of CIS43 can protect against malaria infection for up to 9 months. Here, we developed an in silico pipeline to improve the potency of CIS43 antibody variants by optimizing the binding energy to its target antigen PfCSP. Designed variants showed increased affinity and superior protective efficacy to an in vivo mouse challenge model. The best-designed variant, antibody P3-43, showed ~10-fold improvement in protection relative to CIS43. This novel in silico pipeline provides a generally applicable approach to improve antibody functionality and lead to novel variants to be used for passive prevention against malaria.