Combinatorial pipeline for discovery of anti-GPCR Nanobodies

Biocognon Llc, Pittsburgh PA

Investigators

Abstract

Antibody-based therapeutics are a $200 billion market, and is growing rapidly. Growth will be driven by the antibody discovery sector fueling the demand and innovation. This underscores the need for advanced antibody discovery platforms that efficiently can generate high-quality immune reagents. Existing pipelines encounter substantial screening limitations: (1) Heavy dependence on purified proteins limits access to key antigens of great interest to the industry; (2) The utilization of full antigens impedes targeted domain-specific strategies; (3) Single-antigen screening prevails, hindering simultaneous assessment of multiple antigens in a combinatorial screening; (4) Current platforms lack precise synthetic biology for comprehensive antibody and target antigen design, hindering advanced therapeutic immunoreagent development by not integrating bioinformatics and AI effectively. Biocognon's nanobody (NB) discovery pipeline leverages an advanced self-reporƟng yeast dual biosensor technology, overcoming current field limitaƟons. This innovaƟon signifies a breakthrough in NB screening methodology and in the future applicaƟons of these immunoreagents. Our system enables a 'many-on-many' combinatorial NB screening mode, without the need for physically purifying target anƟgen or anƟbody and can uƟlize targeted domain- specific strategies. As such, our pipeline possesses the precision to target the extracellular domains (loops and NH2- termini) of G-protein coupled receptors (GPCRs). G protein coupled receptors (GPCRs) are high-value therapeuƟc targets represenƟng around 30% of current medicines. However, they are under-represented among biologic drugs, in part because current discovery methods are poorly suited to address these small mulƟ-pass membrane proteins effecƟvely. We believe that Biocognon's innovaƟve technology provides a superior means to discover bioacƟve nanobodies against human GPCRs. Discoveries are expected to lead to development and commercializaƟon of new therapeuƟc treatments for mulƟple human diseases and condiƟons. In this project, we will demonstrate our innovaƟve combinatorial nanobody discovery platiorm for GPCRs uƟlizing our yeast dual biosensor technology through three sequenƟal steps. (1) We will develop a robust 4-piece Golden Gate assembly system, uƟlizing our validated yeast secrete and display vectors as a foundaƟon to facilitate efficient assembly of GPCR/nanobody libraries with high complexity. (2) We will build a fully syntheƟc nanobody library system for the human GPCRs ADRB1 and ADRB2 and conduct single screens to idenƟfy novel nanobodies to these GPCRs and validate their therapeuƟc efficacy. (3) We will Implement a combinatorial screen against four GPCRs (ADRB1, ADRB2, CXCR1, CXCR2) from two subfamilies to demonstrate a fully funcƟonal combinatorial nanobody discovery pipeline. If successful, we will establish a robust foundaƟon to effecƟvely target the complete spectrum of the 400 druggable human non-olfactory GPCRs. This innovaƟon in NB discovery is expected to lead to development and commercializaƟon of new therapeuƟc treatments for mulƟple human diseases and condiƟons.