测试用于成像质谱细胞术的纳米颗粒试剂。
Testing a Nanoparticle Reagent for Imaging Mass Cytometry.
发表日期：2024 Aug 27
作者：
Mahtab Abtahi, Ladan Gheiratmand, Anuroopa Dinesh, Yang Liu, Edmond C N Wong, Hyungjun Cho, Daniel Majonis, Hartland W Jackson, Miralem Mrkonjic, Mitchell A Winnik
来源：
BIOMACROMOLECULES
摘要：
质谱流式细胞仪 (MC) 是一种强大的单细胞分析技术，但在检测低丰度生物标志物方面存在局限性。纳米颗粒 (NP) 试剂通过携带大量重金属同位素提供了增强灵敏度的潜力。在这里，我们报告了基于 NaYF4:Yb3 /Er3 NP 的 NP 报告基因，用于成像质量流式细胞仪 (IMC)。两步配体交换用于用甲氧基-PEG2K-neridronate (PEG-Ner) 和/或聚(磺基甜菜碱甲基丙烯酸酯)-neridronate (PSBMA-Ner) 涂覆 NP 表面。两种修饰都提供了 PBS 缓冲液中的长期胶体稳定性。对扁桃体组织的 IMC 测量表明，与单独的 PEG-Ner 不同，PSBMA-Ner 或 PSBMA-Ner PEG-Ner 的 1:1 混合物在 2 × 1010 NPs/mL 下有效抑制非特异性结合 (NSB)。然而，乳腺癌组织样本显示，NSB 滴度高于 2 × 1010 NPs/mL 时有所增加。使用混合 PEG-Ner 和 PSBMA-Ner 涂层减少 NSB，为使用异双功能 PEG 开发具有生物亲和剂的 NP 缀合物打开了大门，从而实现更灵敏和更特异的 MC 分析。
Mass cytometry (MC), a powerful single-cell analysis technique, has limitations in detecting low-abundance biomarkers. Nanoparticle (NP) reagents offer the potential for enhancing sensitivity by carrying large numbers of heavy metal isotopes. Here, we report NP reporters for imaging mass cytometry (IMC) based on NaYF4:Yb3+/Er3+ NPs. A two-step ligand exchange was used to coat NP surfaces with either methoxy-PEG2K-neridronate (PEG-Ner) and/or poly(sulfobetaine methacrylate)-neridronate (PSBMA-Ner). Both modifications provided long-term colloidal stability in PBS buffer. IMC measurements on tonsil tissue showed that PSBMA-Ner or a 1:1 mixture of PSBMA-Ner + PEG-Ner effectively suppressed nonspecific binding (NSB) at 2 × 1010 NPs/mL, unlike PEG-Ner alone. However, breast cancer tissue samples showed increased NSB at titers above 2 × 1010 NPs/mL. Reduced NSB with mixed PEG-Ner and PSBMA-Ner coatings opens the door for using heterobifunctional PEGs for the development of NP conjugates with bioaffinity agents, enabling more sensitive and specific MC analyses.