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Updated: Aug 4, 2025
Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
Published on: January 17, 2015
Concerted Antibody and Antigen Discovery by Differential Whole-cell Phage Display Selections and Multi-omic Target
Matthew G Cyr1, Henry D Wilson2, Anna-Lena Spierling3
1Skaggs Graduate School of Chemical and Biological Sciences, The Scripps Research Institute, Jupiter, FL, USA; Department of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology, University of Florida, Jupiter, FL, USA. Electronic address: https://twitter.com/CyrialDilutions.
Abstract:
Monoclonal antibody (mAb)-based biologics are well established treatments of cancer. Antibody discovery campaigns are typically directed at a single target of interest, which inherently limits the possibility of uncovering novel antibody specificities or functionalities. Here, we present a target-unbiased approach for antibody discovery that relies on generating mAbs against native target cell surfaces via phage display. This method combines a previously reported method for improved whole-cell phage display selections with next-generation sequencing analysis to efficiently identify mAbs with the desired target cell reactivity. Applying this method to multiple myeloma cells yielded a panel of >50 mAbs with unique sequences and diverse reactivities. To uncover the identities of the cognate antigens recognized by this panel, representative mAbs from each unique reactivity cluster were used in a multi-omic target deconvolution approach. From this, we identified and validated three cell surface antigens: PTPRG, ICAM1, and CADM1. PTPRG and CADM1 remain largely unstudied in the context of multiple myeloma, which could warrant further investigation into their potential as therapeutic targets. These results highlight the utility of optimized whole-cell phage display selection methods and could motivate further interest in target-unbiased antibody discovery workflows.
Insights
This study introduces a novel, target-unbiased antibody discovery method using whole-cell phage display. This approach identified new multiple myeloma cell targets, PTPRG, ICAM1, and CADM1, for potential cancer therapies.
Area of Science:
- Biotechnology
- Immunology
- Oncology
Background:
- Monoclonal antibody (mAb)-based biologics are established cancer treatments.
- Traditional antibody discovery targets a single antigen, limiting the identification of novel specificities.
- There is a need for unbiased methods to discover antibodies against complex cell surface targets.
Purpose of the Study:
- To develop and validate a target-unbiased antibody discovery approach using whole-cell phage display.
- To identify novel antibody specificities and their cognate antigens on multiple myeloma cells.
- To explore new therapeutic targets for multiple myeloma.
Main Methods:
- Generating monoclonal antibodies (mAbs) against native target cell surfaces via phage display.
- Employing improved whole-cell phage display selections combined with next-generation sequencing.
- Utilizing a multi-omic target deconvolution approach to identify antigen identities.
Main Results:
- Successfully generated a panel of over 50 unique mAbs with diverse reactivities against multiple myeloma cells.
- Identified and validated three cell surface antigens: Protein Tyrosine Phosphatase Receptor type G (PTPRG), Intercellular Adhesion Molecule 1 (ICAM1), and Cell Adhesion Molecule 1 (CADM1).
- PTPRG and CADM1 were identified as largely unstudied in multiple myeloma, suggesting potential therapeutic relevance.
Conclusions:
- Optimized whole-cell phage display selection is effective for target-unbiased antibody discovery.
- The identified antigens (PTPRG, ICAM1, CADM1) represent promising new targets for multiple myeloma therapy.
- This unbiased discovery workflow can uncover novel antibody functionalities and therapeutic strategies.
