PDF(Pubmed)
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.
摘要：
基于单克隆抗体(mAb)的生物制品是已建立的癌症治疗方法。抗体发现活动通常针对感兴趣的单个靶标，这固有地限制了揭示新的抗体特异性或功能性的可能性。这里,我们提出了一种用于抗体发现的靶标无偏倚方法，该方法依赖于通过噬菌体展示针对天然靶细胞表面产生mAb。该方法将先前报道的用于改进的全细胞噬菌体展示选择的方法与下一代测序分析相结合,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一组>50mAb,其具有独特的序列和不同的反应性。为了揭示这个小组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用于多体目标反卷积方法。由此,我们鉴定并验证了三种细胞表面抗原：PTPRG，ICAM1和CADM1。PTPRG和CADM1在多发性骨髓瘤的背景下仍未被研究，这可能需要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，并且可以激发对靶标无偏抗体发现工作流程的进一步兴趣。
