Probody therapeutics (PbTx) are engineered with peptide masks that limit their activity and toxicity in healthy tissue. Protease activity in the tumor microenvironment (TME) removes the mask and activates the PbTx in the TME. Kinetic and mutagenesis studies of peptide masks and PbTx are providing insights into how a mask engages a PbTx and blocks its activity in healthy tissue while permitting rapid activation after protease cleavage in the TME.

One of the remaining issues of anti-tumor antibody therapeutics is on-target off-tumor toxicity induced by binding to target antigens expressed in normal tissues. To overcome this issue, we have established a novel antibody technology, called Switch-Ig, which binds to antigens in response to extracellular ATP accumulated higher in tumor microenvironment. We will describe the generation of such environment-responsive antibodies with focused library approach.

Novel technology solutions are needed to overcome the challenges of today’s drug discovery and development. In particular, tailored solutions beyond antibody-antigen binding affinity criteria are required for the discovery of therapeutic antibodies with challenging Target Product Profiles (TPPs). AbCheck’s customized microfluidics solutions address these challenges by meeting the key requirements for potent, function-specific antibodies and enabling functional screening of millions of single cells/day.

Cartilaginous fishes (sharks, skates, rays, and chimera) diverged from the common ancestor with other vertebrates over 450 million years ago and are the oldest lineage to possess immunoglobulins. Among their immunoglobulin repertoire sharks, like camelids, have a heavy chain only isotype (IgNAR) with small, soluble, single-domain variable regions (VNARs). We will discuss the generation, selection, and downstream engineering of VNARs for development as diagnostic and therapeutic agents.

The unique characteristics of heavy chain-only antibodies (HCAbs) endow these molecules with the potential to target novel epitopes and reduce the complexity of downstream engineering. We have adapted our yeast-based platform to allow for the selection of high-affinity, target-specific HCAbs from immunized camelids. Using this methodology, we have discovered and optimized HCAbs against a diverse range of antigens, including multi-spanning membrane proteins and other difficult targets.

Pioneer is a new phage display Fab antibody library for the selection of therapeutic antibodies. The design incorporates Bio-Rad’s extensive experience of phage display to create a superior library. Pioneer is the largest functional Fab antibody library available, and incorporates a novel selection principle. The design, features, and data will be presented for the first time, including SpyDisplay, the new selection system based on SpyTag protein ligation technology.

Loop Genomics has developed a synthetic long read sequencing technology that leverages existing short read sequencers to enable highly accurate single-molecule long-read sequencing on any short-read sequencer. In this talk we will explore LoopSeq and how it is applied to provide additional, previously inaccessible layers of information from short-read sequencers for a wide variety of applications such as phage display, single B-cell discovery and more. 

Miniproteins are a powerful yet underutilized therapeutic modality. They are only 30-90 amino acids in length, yet they adopt a folded tertiary structure like a much larger protein; this structure enables miniproteins to bind with high affinity and specificity to their targets. Miniproteins found in nature are very challenging to engineer to bind new targets. We solved this problem using computational de novo design, finally unlocking miniproteins for therapeutic development.

We have developed a chemical biology platform to engineer protein-small molecule hybrids. Combinatorial libraries of protein variants are displayed on yeast, chemically conjugated with pharmacophore, and sorted for strong, selective binders. We engineered hybrids that exhibit superior enzymatic inhibitory potency and selectivity relative to either protein or pharmacophore alone. Hybrids were engineered with different conjugation sites, linkers, protein sequences, and targets, which demonstrates the breadth of the approach.

Twist Biopharma, a division of Twist Bioscience, combines high-throughput DNA synthesis technology with deep expertise in antibody engineering to provide end-to-end antibody discovery solutions. The result is a make-test cycle that yields better antibodies against more diverse and challenging targets. Twist Biopharma will continue to optimize and expand its library synthesis and screening capabilities in partnership with other discovery technologies to further utilize the Twist make-test cycle. 

Medication non-adherence (non-compliance) represents a major barrier to effective clinical care. In developed nations, only 50% of patients take their medications as prescribed. In his seminar, Dr. Traverso will present a series of novel technologies being developed with the goal to enhance and facilitate medication administration. Specifically, Dr. Traverso will discuss the development of new technologies for the delivery of macromolecules through the oral route.

Novel peptides that mimic IL-2 and IL-7 in a variety of cell-based assays and demonstrate desired activity in animal models will be described. The peptides have been successfully incorporated as components of bispecific constructs to add cellular selectivity through cis-activation or to incorporate multiple cytokine activities in a single molecule. Being unrelated in sequence to the natural cytokine, a major advantage is the avoidance of ADAs that neutralize endogenous cytokine.

Cytokines control all immune cell activities. Cytokines dimerize/oligomerize surface receptors to activate signalling. Given the strong crosstalk and shared usage of key components of their signalling pathways, a long-standing question in the field pertains to how functional diversity is achieved by cytokines. I will present recent work from my laboratory addressing how cytokine-receptor binding parameters modify cytokine responses and how this helped to design less toxic cytokine-based therapies.

To promote immune activation within tumors, we chemically conjugated a TLR9 agonist to a unique engineered integrin-binding peptide that localizes to various types of solid tumors. Intravenous dosing of this tumor-targeted immunostimulant transforms the immunosuppressive tumor microenvironment into one abundant with activated lymphocytes and results in robust T cell-mediated tumor regression, and in some cases cures, in murine breast and pancreatic cancer models.

We are interested in building genetic systems that have extremely high mutation rates in order to 1) speed up and scale the evolution of proteins, enzymes, and antibodies in vivo and 2) record transient information, such as lineage relationships or exposure to biological stimuli, as durable genetic information in situ. I will discuss our work on OrthoRep, a highly error-prone orthogonal DNA replication system that drives the rapid continuous mutation and evolution of user-selected genes. I will focus on OrthoRep’s application to yeast-display antibody evolution and comment on the value of depth and scale in evolutionary search.

I will provide an introduction to my strategic vision, Biopharmaceutical Informatics, and describe how it can be used to improve the efficiency of biologic drug discovery and development cycles.

Higher affinity antibodies may expand the druggable target space and be more efficacious, particularly as agonists or antagonists. Attobodies are bi-epitopic nanobodies using proprietary engineering for superior potency and specificity. We routinely deliver Attobodies with low picomolar affinity without affinity maturation, reducing development time by 3 – 6 months and avoiding off-target risk. Attobodies are small and stable, easily humanized, and easily modified, e.g., Fc fusion, bispecific configuration.

The Specifica Generation3 Library Platform is based on highly developable clinical scaffolds, into which natural CDRs purged of sequence liabilities have been embedded. The platform directly yields highly diverse, high affinity (20% subnanomolar), developable (>80% lack biophysical liabilities), drug-like antibodies as potent as those from immune sources. This talk will discuss the Generation3 concept and its application to antibodies and VHH scaffolds of clinical interest.