Antibodies are complex proteins naturally generated by the immune system to neutralize foreign pathogens such as bacteria or viruses. B cells, a white blood cell type responsible for the generation of antibodies in response to pathogens, secrete billions of antibodies with different specificities into the bloodstream. Antibodies are structurally distinct Y-shaped proteins formed through the combination of two long proteins, called heavy chains, and two short proteins, called light chains. Each heavy and light chain pair forms a binding site where the antibody specifically binds its target, otherwise known as an antigen, at the Fab domain of the antibody molecule.
The specificity of each antibody to a target, and the potency of its binding strength to that target are defined by the amino acid sequences of heavy and light chains in the Fab domain of the antibody molecule. The other end of the antibody, called the Fc domain, is responsible for communication between the antibody and the rest of the immune system. Fc domains bind to various receptors and cause immune system effector responses.
Therapeutic antibodies are typically non-naturally occurring, or recombinant, antibodies specifically developed to treat human diseases by binding to certain proteins, and thereby modulating key biological processes. Therapeutic antibodies are injectable products that are typically dosed subcutaneously or intravenously.
Our innovative platform is designed to replicate the natural process of somatic hypermutation (SHM) embedded within the human immune system to rapidly develop a diverse range of therapeutic-grade antibodies in vitro. SHM is a critical, endogenous process that generates the essential antibody diversity required to develop a natural immune response to pathogens. Our genomes encode a limited number of antibody genes, which are insufficient to generate antibodies against the wide variety of foreign pathogens encountered from the external environment. SHM enables our immune system to expand the limited diversity encoded within our genomes to the billions of antibody specificities required to defend ourselves against external pathogens.
The key enzyme required for SHM is called activation-induced cytidine deaminase, or AID. AID has been genetically conserved throughout mammalian biology and is required for the non-random mutagenesis pattern associated with SHM. AID is specifically expressed by B cells after contact with a foreign pathogen and modifies antibody sequences in a non-random fashion. Through SHM, B cells evolve antibodies with the potency and specificity required to clear the foreign pathogen. However, within the in vivo environment, SHM does not generally progress to the creation of high potency antibodies or develop antibodies against the body’s own proteins.
By coupling in vitro SHM with our mammalian cell system that simultaneously displays and secretes antibodies, we believe our SHM platform is able to rapidly identify and mature antibodies with desired functional activity to high potency while simultaneously mitigating the risks associated with manufacturing. We introduce AID into mammalian cells to replicate the non-random mutagenesis SHM pattern observed within B cells in vivo. Starting with a library of either fully-human or humanized antibodies, our platform generates AID-based variants of the starting antibody library throughout the process. We have demonstrated that the pattern of mutagenesis we observe in vitro using our platform technology closely mimics the pattern observed among in vivo generated antibodies, thereby increasing confidence that antibodies generated by our platform will be tolerated when used as therapeutic drugs in humans.
By selecting antibodies based on their antigen binding from the broad antibody library population our SHM platform develops, we are able to evolve in an iterative fashion the binding potency and function of antibodies to levels that we believe will be required for therapeutic use. We believe this approach allows us to rapidly generate antibodies with high binding potency against a target. Through this approach, we have successfully generated therapeutic antibody product candidates to more than 25 targets, including targets that have been challenging for competing antibody technology platforms.
Each evolving antibody is expressed within the SHM-active mammalian cell to concurrently (i) display the evolved antibody on the cell surface to permit cell sorting selection for potency properties while (ii) the same antibody is secreted into the extracellular media at sufficient quantities to permit functional assays to be conducted. In this manner, the evolving antibodies expressed by each transfected cell are assessed in a high-throughput fashion for the desired functional activity relevant to the therapeutic mechanism.
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Diversity Against Difficult Targets
We are able to generate an unprecedented diversity of antibodies by applying SHM-based diversification outside of the constraints of an in vivo environment. This enables us to develop antibodies against human targets that we believe have not otherwise been accessible to prior technologies.
Because our platform generates highly-potent antibodies, we are potentially able to modulate every extracellular target associated with human disease, and believe only small therapeutic doses may be required to mediate therapeutic effect in vivo.
Functional Activity Selection
Our mammalian cell system simultaneously displays and secretes antibodies during the antibody discovery process, allowing us to incorporate functional assays throughout the process and focus on producing product candidates that are optimized for the desired therapeutic activity.
Our platform technology enables us to generate therapeutic-grade antibodies and initiate subsequent preclinical manufacturing and toxicology studies.
By utilizing our mammalian cell display system, we believe our approach increases the probability of success in manufacturing and commercialization by mitigating risks associated with antibody expression, formulation and stability.
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Martin E. Dahl, Eric Hare, Kenneth Luu, Bruce Randazzo and Paul Lizzul
Taku Okazaki and Tasuku Honjo
21st Century Center of Excellence Formation and Department of Immunology and Genomic Medicine, Graduate School of
Medicine, Kyoto University, Yoshida-Konoe, Sakyo-ku, Kyoto 606-8501, Japan
Marilyn R. Kehry, Gerald Manorek, Janean Fisher, Natasha Del Cid, Allison Rooks, Laurence Altobell III, Gregory Gold, Morena Shaw, Robert Morse, Margaret Marino, Jessie-Farah Fecteau, and Stephen Parmley
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Caroline Stienne, Richard Virgen-Slane, Lisa Elme´ n, …, Kenneth M. Murphy,
Carl F. Ware, John R. Sedy
ANB032, a Novel BTLA/HVEM Therapeutic Antibody, Inhibited T cells Derived from Atopic Dermatitis Patients In-vitro
Matthew Hsu, Stephanie Baguley, Tyson Chase, Justin Choi, Jennifer Michaels, Safak Yalcin, Eric Hare, Gerald Manorek, April Fraley, Gregory Gold, Robert Morse, Margaret Marino, JingUeiVerkade, Stephen Parmley and Martin Dahl.
Soluble BTLA (sBTLA) is Induced Following Targeting of BTLA In-vivo by ANB032, a Novel BTLA/HVEM Modulator Therapeutic Antibody for The Treatment of Autoimmune Disease
Martin E. Dahl, Tyson Chase, Stephanie Baguley, Eric Hare, Gerald Manorek, Kristine Storey, April Fraley, Gregory Gold, Robert Morse, Margaret Marino, Scott Lashbrook, Justin Choi, JingUei Verkade, Yu-yu Ren and Stephen Parmley,
Discovery and Characterization of ANB032, a Novel BTLA/HVEM Checkpoint Modulator for Autoimmune/Inflammatory Disease
Martin E. Dahl, Jean da Silva, Gerald Manorek, Natasha Del Cid, April Fraley, Gregory Gold, Robert Morse, Margaret Marino, JingUei Verkade, Yu-yu Ren, Janean Young, Paul Fisher,
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A Phase 1 Study of ANB019, an Anti-Interleukin-36-Receptor (IL-36R) Monoclonal Antibody, in Healthy Volunteers
Khanskaya et al. 2018 May. Poster presented at the 2018 European Academy of Allergy and Clinical Immunology (EAACI) Congress.
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Laken et al. Poster presented at EORTC-NCI-AACR Symposium on Molecular Targets and Cancer Therapeutics 2016.
Kehry et al. Poster presented at AACR Annual Meeting 2015.
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Jun et al. Poster presented at AACR Annual Meeting 2014.