A wide-pore silica-based superficially porous material with a high coverage phenyl bonding was evaluated for the analysis of monoclonal antibodies and antibody-drug conjugates. This new material is based on 2.7 μm particles having a shell thickness of 0.40 μm and average pore size of approximately 450 Å.

Various important features of this reversed phase column technology were explored, including kinetic performance for large biomolecules (i.e. speed of analysis, efficiency and peak capacity), recovery of proteins, selectivity for resolving modifications, and the possibility to reduce the amount of trifluoroacetic acid in the mobile phase. A systematic comparison was also performed with other existing modern wide-pore phases possessing differences in structure/morphology and chemistry.

If all these figures of merit are considered, it is clear that this phenyl bonded wide-pore superficially porous stationary phase is one of the most promising materials to have been developed in recent years. Indeed, it offers kinetic performance comparable to the most efficient wide-pore SPP column on the market. In terms of protein recovery, this new phase was found to be superior to silica-based and silica-hybrid C4 bonded materials, particularly with separations performed at sub-80 °C temperature. Under such conditions, it in fact shows recoveries that are quite similar to a divinyl benzene (DVB) polymer-based material. More importantly, due to its unique, high coverage phenyl bonding, it offers additional steric effects and potentially even π-π interactions that yield advantageous selectivity for mAb sub-unit peaks and ADC species as compared to commonly used C4 or C18 bonded phases. Last but not least, mobile phases consisting of only 0.02–0.05% trifluoroacetic acid can be successfully used with this column, without significant loss in recovery and peak capacity.

评估了具有高覆盖率的苯基键合的宽孔二氧化硅基表面多孔材料，用于分析单克隆抗体和抗体-药物偶联物。这种新材料基于2.7μm颗粒，壳厚度为0.40μm，平均孔径约为450Å。

探索了这种反相柱技术的各种重要特征，包括大型生物分子的动力学性能（即分析速度，效率和峰容量），蛋白质回收率，解析修饰的选择性以及减少三氟乙酸含量的可能性。流动相。还对其他在结构/形态和化学方面存在差异的现代宽孔相进行了系统的比较。

如果考虑所有这些优点，很显然，这种苯基键合的宽孔表面多孔固定相是近年来开发的最有前途的材料之一。实际上，它提供的动力学性能可与市场上最有效的宽孔SPP色谱柱相媲美。在蛋白质回收方面，发现该新相优于硅胶基和二氧化硅杂化的C4键合材料，尤其是在低于80°C的温度下进行分离时。在这样的条件下，它实际上显示出与基于二乙烯基苯（DVB）聚合物的材料非常相似的回收率。更重要的是，由于其独特的高覆盖苯基键，与常用的C4或C18键合相相比，它提供了额外的空间效应以及潜在的π-π相互作用，从而对mAb亚基峰和ADC物种产生了有利的选择性。最后但并非最不重要的一点是，仅由0.02–0.05％的三氟乙酸组成的流动相可成功用于此色谱柱，而回收率和峰容量却没有明显损失。