A CHO Cell-Based Surface Display Platform for Comprehensive Serological Profiling

Building a CHO cell surface display library of all human proteins to enable comprehensive, high-throughput autoantibody profiling across autoimmune diseases and vaccine responses.
Completed Other Autoantibodies Autoantigens Biological & Mechanistic Biomarker Discovery Experimental Platforms & Models Functional Genomics & CRISPR Immune Profiling In Vitro Models Translational & Clinical

Funding Year: 2023

Project Overview

Autoantibody production is a hallmark of autoimmune disease, and the amount, affinity, and type of autoantibodies vary across disease stages and conditions — making comprehensive profiling a powerful tool for diagnosis and therapeutic targeting. This project constructs a mammalian cell library displaying potentially all human proteins in their natural conformation and with post-translational modifications, using ExpiCHO cells and CRISPR-Cas9-based knock-in technology. Unlike prior platforms limited to soluble proteins, this system can display membrane-bound cytokines, ion channels, and multimeric proteins. Key advances include confirmed surface expression of 175 autoantigens and 123 cytokines, single-copy transgene insertion for uniform expression, and high recovery rates (>98%) from liquid nitrogen storage — enabling direct use in autoantibody screening without further culturing.

Impact & Innovation

A universal platform for reading the autoantibody landscape.

 

By displaying all human proteins on CHO cell surfaces in native conformation — including membrane-bound and multimeric proteins that prior platforms cannot capture — this project removes a foundational barrier to comprehensive autoimmune serological profiling.

  • Overcomes the major limitation of existing autoantibody profiling platforms by displaying membrane-bound cytokines, ion channels, and multimeric proteins in native conformation with post-translational modifications intact
  • Establishes a high-throughput, quantitatively accurate serological screening platform with direct applications in autoimmune diagnosis, cancer therapy monitoring, and vaccine response evaluation
  • Advances the Consortium’s Integrated Data and Discovery Platforms pillar by creating a broadly shareable, reusable cell library that enables comprehensive autoantibody profiling across autoimmune conditions and research settings
Research Approach

A framework designed for discovery

This project applies CRISPR-Cas9-based knock-in technology to construct a comprehensive CHO cell surface display library of human proteins for high-throughput autoantibody profiling, combining cell engineering with functional validation to establish a broadly applicable serological screening platform.

CRISPR-Cas9/HDR-mediated single-copy transgene insertion into a single chromatin locus in ExpiCHO cells for uniform gene expression; surface display validation of 175 autoantigens and 123 cytokines confirming membrane expression and functional retention; high-density culture optimization to achieve at least 500-fold coverage of all human protein-coding genes; and liquid nitrogen storage recovery testing confirming >98% cell viability for direct screening use.

ExpiCHO cell surface expression datasets for 175 autoantigens and 123 cytokines; CRISPR-Cas9 transgene integration and expression uniformity datasets; cell density and viability datasets validating library coverage and storage recovery; and functional assay data confirming membrane-bound protein retention of natural conformation and post-translational modifications.

Establishment and validation of a comprehensive CHO cell surface display library enabling high-throughput, quantitatively accurate autoantibody profiling across autoimmune conditions, cancer therapy, and vaccine response evaluation. The platform addresses the critical limitation of prior technologies by displaying the full range of human protein types, and is designed for broad research and clinical diagnostic applications.

Investigators & Institutions

Powering the science

Principal Investigator

Leyuan Ma, PhD, Colton Consortium Member

Assistant Professor, Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania

Colton Centers & Institutions

Colton Center for Autoimmunity at the University of Pennsylvania

Perelman School of Medicine