Concerted Antibody and Antigen Discovery by Differential Whole-cell Phage Display Selections and Multi-omic Target Deconvolution. Issue 10 (15th May 2023)

• Record Type: Journal Article
• Authors: Cyr, Matthew G.; Wilson, Henry D.; Spierling, Anna-Lena; Chang, Jing; Peng, Haiyong; Steinberger, Peter; Rader, Christoph
• Is Part Of: Journal of molecular biology. Volume 435:Issue 10(2023)
• Publication Date: 2023-05-15
• Subjects: Fab -- multiple myeloma -- PTPRG -- ICAM1 -- CADM1; aa amino acid -- BSA bovine serum albumin -- CADM1 cell adhesion molecule 1 -- CAR-T chimeric antigen receptor T cell -- DMEM Dulbecco's Modified Eagle's Medium -- DPBS Dulbecco's PBS -- ECD extracellular domain -- FACS fluorescence-activated cell sorting -- FBC Fab-phage biotinylation and capture -- FBS fetal bovine serum -- FDA Food and Drug Administration -- GARS glycyl-tRNA synthetase -- HCDR3 heavy chain complementarity determining region 3 -- huCH1 human constant heavy chain domain 1 -- huCL human constant light chain domain -- ICAM1 intercellular adhesion molecule 1 -- IMAC immobilized metal affinity chromatography -- IP immunoprecipitation -- LCDR3 light chain complementarity determining region 3 -- LC-MS/MS liquid chromatography tandem mass spectrometry -- mAb monoclonal antibody -- MM multiple myeloma -- NGS next-generation sequencing -- NT non-target -- PBMCs peripheral blood mononuclear cells -- PBS phosphate-buffered saline -- PS penicillin–streptomycin -- PTM post-translational modification -- PTPRG protein tyrosine phosphatase receptor type G -- rbVH rabbit variable heavy chain domain -- rbVL rabbit variable light chain domain -- RT room temperature -- SCLC small cell lung cancer -- SPR surface plasmon resonance -- TBS Tris-buffered saline -- TCR T cell receptor -- WCP whole-cell panning; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2023.168085
• Languages: English
• ISSNs: 0022-2836

Abstract

Graphical abstract: Highlights: There is a need for new cell surface antigens in antibody-based cancer therapy. Target-agnostic Fab-phage library selection and sequencing unveil new antigens. Combining proteomics and transcriptomics robustly facilitates antigen identification. Method provides a streamlined approach to antibody drug and target discovery. 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.