The addition of supercharging (SC) reagents in electrospray ionization coupled mass spectrometry (ESI-MS) has demonstrated several advantages for protein analysis such as reduced required mass range of the instrument, narrowed charge-state distribution, increased sensitivity, and adduct suppression. The potential use of SC reagents to improve analyses of larger and complex protein molecules such as monoclonal antibodies and antibody–drug conjugates (ADCs) has not been previously reported. In this study, the effect of seven SC reagents (meta-nitrobenzyl alcohol (m-NBA), dimethyl sulfoxide (DMSO), ortho-nitroanisole (o-NA), propane sultone (PS), ethylene carbonate (EC), propylene carbonate (PC), and sulfolane) on ESI-MS acquired spectra of deglycosylated, intact, and reduced trastuzumab and a vcMMAE–trastuzumab ADC was investigated under denaturing conditions. The addition of any of the SC reagents resulted in a higher average charge state observed for all tested reagents for both trastuzumab and the ADC and a narrower charge-state envelope for o-NA and 1% sulfolane for trastuzumab. However, improved peak shapes or increased sensitivity was observed for several reagents, overall increasing the spectra quality. Finally, it was shown that SC reagents can be safely used for ADC analysis without impacting the obtained drug-to-antibody (DAR) values, as all DAR values were within 0.1 from the control sample.

中文翻译：

---

增压剂在改进单克隆抗体和抗体-药物偶联物质谱分析中的潜在用途

在电喷雾电离耦合质谱 (ESI-MS) 中添加增压 (SC) 试剂已经证明了蛋白质分析的几个优点，例如减少仪器所需的质量范围、缩小电荷态分布、提高灵敏度和加合物抑制。以前没有报道过使用 SC 试剂来改进更大和复杂的蛋白质分子（如单克隆抗体和抗体-药物偶联物 (ADC)）的分析。在本研究中，七种 SC 试剂（间硝基苯甲醇（m -NBA）、二甲基亚砜（DMSO）、邻硝基苯甲醚（o-NA)、丙磺酸内酯 (PS)、碳酸亚乙酯 (EC)、碳酸亚丙酯 (PC) 和环丁砜) 在 ESI-MS 上获得的去糖基化、完整和还原曲妥珠单抗的光谱以及在变性条件下研究 vcMMAE-曲妥珠单抗 ADC . 添加任何 SC 试剂导致观察到的所有测试试剂对于曲妥珠单抗和 ADC 的平均电荷状态更高，并且对于曲妥珠单抗，o - NA 和 1% 环丁砜的电荷状态包络更窄。然而，观察到几种试剂的峰形改善或灵敏度增加，从而总体上提高了光谱质量。最后，结果表明 SC 试剂可以安全地用于 ADC 分析，而不会影响获得的药物抗体 (DAR) 值，因为所有 DAR 值都在控制样本的 0.1 范围内。