Skaggs 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.
Skaggs Graduate School of Chemical and Biological Sciences, The Scripps Research Institute, Jupiter, FL, USA.
J Mol Biol. 2023 May 15;435(10):168085. doi: 10.1016/j.jmb.2023.168085. Epub 2023 Apr 3.
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.
单克隆抗体 (mAb)- 为基础的生物制剂是癌症治疗的成熟方法。抗体发现通常针对单一感兴趣的靶标，这从根本上限制了发现新的抗体特异性或功能的可能性。在这里，我们提出了一种针对抗体发现的无目标偏向方法，该方法依赖于通过噬菌体展示生成针对天然靶细胞表面的 mAb。该方法结合了先前报道的用于改进全细胞噬菌体展示选择的方法和下一代测序分析，以有效地鉴定具有所需靶细胞反应性的 mAb。将该方法应用于多发性骨髓瘤细胞，产生了超过 50 种具有独特序列和多样化反应性的 mAb 。为了揭示该面板识别的同源抗原的身份，来自每个独特反应性簇的代表性 mAb 被用于多组学靶标去卷积方法。由此，我们鉴定并验证了三个细胞表面抗原：PTPRG、ICAM1 和 CADM1。PTPRG 和 CADM1 在多发性骨髓瘤的背景下仍然在很大程度上未被研究，这可能值得进一步研究它们作为治疗靶点的潜力。这些结果突出了优化的全细胞噬菌体展示选择方法的实用性，并可能激发人们对无目标偏向的抗体发现工作流程的进一步兴趣。
