This article reported a new strategy for resin activation with divinyl sulfone using catalytic oxa-Michael addition in a controllable manner. By screening a variety of organocatalysts, PPh₃ and DMAP stand out with high catalytic efficiency in aprotic solutions. X-ray photoelectron spectroscopy (XPS) analysis indicates high reaction efficiency and less side reactions than traditional aqueous reactions, resulting in high activation density. A maximum activation density of 157.5 ± 1.2 μmol/g resin was achieved in 12 h using PPh₃ as catalyst, which is 1.5 times higher than the traditional aqueous reactions. Followed by conjugation with a chromatographic ligand, i.e., 4-mercaptoethyl pyridine (MEP), the resin is capable of antibody purification. Using IgG and BSA as model proteins, adsorption isotherms and dynamic binding behavior of the resin samples were investigated. A higher affinity and dynamic binding capacity of IgG was observed on resins with higher ligand density. Finally, the resin samples were applied to the purification of a therapeutic monoclonal antibody from cell culture supernatant. The recovery of the resin samples with high ligand density are 70% higher than those of the commercial resin (MEP HyperCel). Moreover, our method achieves a controllable chromatographic ligand density by varying reaction times, which is useful to clarify the density-affinity relationship and improve process-scale antibody purification.

本文报道了一种新的策略，可控制的方式是使用催化的oxa-Michael加成法用二乙烯基砜进行树脂活化。通过筛选各种有机催化剂，PPh ₃和DMAP在非质子传递溶液中具有很高的催化效率。X射线光电子能谱（XPS）分析表明，与传统的水性反应相比，反应效率高，副反应少，因此活化密度高。使用PPh ₃作为催化剂，在12小时内可达到157.5±1.2μmol/ g树脂的最大活化密度，这是传统水性反应的1.5倍。随后与色谱配体结合，即，4-巯基乙基吡啶（MEP），该树脂能够进行抗体纯化。以IgG和BSA为模型蛋白，研究了树脂样品的吸附等温线和动态结合行为。在具有更高配体密度的树脂上观察到了更高的IgG亲和力和动态结合能力。最后，将树脂样品应用于从细胞培养上清液中纯化治疗性单克隆抗体。具有高配体密度的树脂样品的回收率比市售树脂（MEP HyperCel）高70％。而且，我们的方法通过改变反应时间来实现可控的色谱配体密度，这对于阐明密度亲和关系和改善工艺规模的抗体纯化是有用的。