Next-Generation Antibody Discovery Enabled By NativeMP™ and Phage Display Technology

The Significance of P2X4 Receptors

The P2X4 receptor is part of the P2X family of ATP-gated ion channels expressed on vascular endothelial cells, cardiac myocytes, and immune cells, and crucial for cellular signaling and transport pathways. P2X4 receptor’s involvement in such diverse and critical cellular pathways highlights its potential as a therapeutic target. It has emerged as a prime target for drug development due to its involvement in critical cardiovascular and inflammatory processes.

Its involvement in such diverse and critical cellular pathways underscores its potential as a therapeutic target. However, effective tools for studying and modulating native P2X4 have been limited. Most existing antibodies fail to bind P2X4 due to the use of irrelevant antigens, such as synthetic peptides, which do not mimic the protein’s native structure.

The anti-P2X4 antibodies developed through the collaboration between ProteoGenix and Cube Biotech address this challenge by specifically targeting full-length native P2X4. These antibodies pave the way for:

• Better understanding of P2X4 biology in health and disease.
• Development of targeted therapies for conditions involving P2X4 dysregulation.
• Improved tools for basic and translational research.

By stabilizing P2X4 in a native-like environment and generating fully human antibodies, this collaboration delivers cutting-edge solutions for researchers and therapeutic developers.

A Synergistic Approach: Cutting-Edge Technologies.

Step-by-Step Technology Overview

• Stabilizing Native P2X4 with Cube Biotech’s NativeMP™ Platform

Cube Biotech’s proprietary NativeMP™ copolymer platform was employed to extract and stabilize the full-length P2X4 receptor directly from the cell membrane, preserving its native structural and functional integrity, critical for identifying antibodies that target the biologically relevant form of P2X4. The stabilized protein underwent a two-step purification process involving Rho-MagBeads for initial extraction and Size Exclusion Chromatography to achieve high purity, followed by biophysical characterization to validate its structure and ensure readiness for downstream antibody screening.

• Naive Antibody Library Screening with Phage Display

ProteoGenix provides an extensive range of naïve antibody libraries tailored for discovering therapeutic antibody discovery using phage display. These libraries offer an excellent basis for identifying high-affinity binders. For this project, ProteoGenix used its LiAb-SFMAX™ human library, a powerful resource for rapid and effective antibody discovery.

The library consists of 368 donors from five different ethnic groups. Its extensive genetic pool ensures broad coverage and the ability to target a wide range of human antigens and provides a large antibody repertoire in both scFv and Fab formats, covering 5.37 x 10¹⁰ unique clones, increasing the likelihood of rapidly identifying high-affinity candidates. And because the antibodies are fully human from the start, no additional humanization steps are required, significantly reducing development time, cost and complexity while accelerating the path to clinical application.

By combining diversity, scale, and versatility, the LiAb-SFMAX™ library delivers a highly efficient solution for antibody discovery projects targeting even the most challenging proteins.

• Recombinant Antibody Production : XtenCHO Expression System

Once the top 2 candidates were identified, ProteoGenix’s XtenCHO™ system facilitated their production. This platform is optimized for high-yield, low-endotoxin output, delivering antibodies with >98% purity at a scalable g/L level, ensuring readiness for research and therapeutic applications.

Unique Advantages of the Anti-P2X4 Antibodies

The collaboration resulted in two high-performing antibody candidates confirmed for their exceptional binding affinity to native P2X4 receptors. These antibodies stand out due to their:

• Fully Human Sequences: Eliminating the need for humanization, saving time and resources in clinical development.
• High Purity and Low Endotoxin Levels: Meeting stringent standards for therapeutic and research use.
• Licensing-Ready Status: With all potential IP-related hurdles addressed, these antibodies are primed for seamless integration into therapeutic pipelines..

Why our anti-P2X4 antibodies could be your next lead candidate?

The collaboration resulted in two high-performing antibody candidates confirmed for their exceptional binding affinity to native P2X4 receptors.

• Fully Human: No Need For Further Humanization
• High Binding Affinity Confirmed To Native Human P2X4
• High Production Yield & High Purity Validated
• Free Of Intellectual Property and available for licensing

A Vision for Drug Discovery and Research

This collaboration showcases the power of combining advanced membrane protein stabilization with state-of-the-art antibody discovery technologies. ProteoGenix and Cube Biotech have successfully removed many of the barriers that have previously hindered P2X4-focused antibody development, offering new hope for therapies targeting cardiovascular and inflammatory diseases.

Interested in Licensing These Sequences? The fully human anti-P2X4 antibodies are now available for licensing. Don’t miss the opportunity to secure these groundbreaking tools for your next therapeutic or research breakthrough